Carrot leaves improve color and xanthophyll content of egg yolk in laying hens but are not as effective as commercially available marigold fortificant1.

Yellow and orange egg yolks are good sources of xanthophyll carotenoids, consumption of which is associated with health benefits, such as cancer prevention, eye health, and bone health. Industrial feed fortificants used to improve egg yolk color and carotenoid concentration typically are derived from marigold flowers. Green leafy vegetables are also concentrated sources of the xanthophylls lutein and zeaxanthin (L+Z), but they have not been rigorously evaluated in laying hen feeds as a yolk colorant. The addition of food manufacturing byproducts, including carrot leaves, to animal feed is a promoted method of improving animal nutrition. The ability of dehydrated carrot leaves to improve egg yolk color and L+Z concentration was evaluated by feeding laying hens (n = 40) white maize-based feeds fortified with 2 different dehydrated carrot leaves, marigold as a positive control, or no fortificant as a negative control for 28 D. After a 7-D washout period, the hens were separated into 4 groups, and eggs were collected every other day. Yolks were analyzed by using a portable colorimeter to define the color space and by ultra-performance liquid chromatography to determine the carotenoid profile. Carotenoid concentration rapidly declined from day 0 to 8, confirming adequate washout conditions. The white maize negative control (WM) day 28 lutein concentration (3.59 ± 0.51 nmol/g) was significantly less than orange-carrot leaf-treated (OCL) (5.34 ± 0.36 nmol/g) and red-carrot leaf-treated hens (RCL) (5.92 ± 1.00 nmol/g) in addition to the marigold-treated hens (MG). However, MG was significantly higher than both leaf-treated groups. From day 8 (3.93 ± 0.74 nmol/g) to 28 (9.32 ± 1.66 nmol/g), MG had the largest increase in lutein and was the only treatment to surpass day 0 initial concentrations (8.50 ± 1.64 nmol/g). A similar trend was observed for zeaxanthin and was reflected in the color space.

State-dependent domicile leaving rates in Anopheles gambiae.

BACKGROUND: Transmission of Plasmodium greatly depends on the foraging behaviour of its mosquito vector (Anopheles spp.). The accessibility of blood hosts and availability of plant sugar (i.e., nectar) sources, together with mosquito energy state, have been shown to modulate blood feeding (and thus biting rates) of anopheline mosquitoes. In this study, the influence of mosquito starvation status and availability of nectar on the decision of female Anopheles gambiae mosquitoes to leave a bed net-protected blood host was examined. METHODS: Two small-scale mesocosm experiments were conducted using female mosquitoes starved for 0, 24 or 48 h, that were released inside a specially constructed hut with mesh-sealed exits and containing a bed net-protected human volunteer. Floral cues were positioned on one side of the hut or the other. Several biologically plausible exponential decay models were developed that characterized the emigration rates of mosquitoes from the huts. These varied from simple random loss to leaving rates dependent upon energy state and time. These model fits were evaluated by examining their fitted parameter estimates and comparing Akaike information criterion. RESULTS: Starved mosquitoes left domiciles at a higher rate than recently fed individuals however, there was no difference between 1- and 2-day-starved mosquitoes. There was also no effect of floral cue placement. The best fitting emigration model was one based on both mosquito energy state and time whereas the worst fitting model was one based on the assumption of constant leaving rates, independent of time and energy state. CONCLUSIONS: The results confirm that mosquito-leaving behaviour is energy-state dependent, and provide some of the first evidence of state-dependent domicile emigration in An. gambiae, which may play a role in malarial transmission dynamics. Employment of simple, first-principle, mechanistic models can be very useful to our understanding of why and how mosquitoes leave domiciles.

Incidence and Severity of Cavity Spot of Carrot as Affected by Pigmentation, Temperature, and Rainfall.

Field trials to determine the effect of carrot pigmentation and weather parameters on cavity spot (CS) of carrot were conducted in the Holland/ Bradford Marsh region of Ontario between 2002 and 2009. In all, 23 colored carrot cultivars from the United States Department of Agriculture (USDA) Agricultural Research Service breeding program at the University of Wisconsin (n = 5) and commercial seed companies (n = 18) were seeded in organic soil (pH 6 to 7, 45 to 75% organic matter) in late May to early June and harvested in late October or early November. Carrot roots were assessed for CS severity midseason and postharvest. Evaluations postharvest indicated that the purple pigmented carrot from breeding line 'USDA 106-3' and cultivars 'Purple Rain' and 'Purple Haze' consistently had low CS severity. The orange-pigmented 'USDA 101-23', 'Cellobunch', 'YaYa', and 'Envy' had moderate CS; and the red-pigmented carrot breeding line 'USDA 104-3' and cultivars 'Atomic Red', 'Proline Red', 'Dragon', and an unnamed line from India had high CS. Differences in CS severity in carrot cultivars between evaluations at midseason and postharvest suggest that some carrot cultivars are more susceptible to Pythium spp. inoculum in soil (alloinfection) and others to secondary infection (autoinfection) that can be attributed to the Pythium sp. involved in CS. CS severity was positively correlated with total rainfall 2 and 3 months after seeding, and was negatively correlated with number of days with air temperature ≥30°C 3 and 4 months after seeding. Soil temperature and total rainfall were the best predictors of CS incidence and severity. These results could allow a forecast of disease incidence and severity at harvest.

Microbiological selenate to selenite conversion for selenium removal.

Removal of the toxic selenium compounds selenite and selenate from waste water before discharge is becoming increasingly imperative in industrialized countries. Bacteria can reduce selenate to selenite, but also further to elemental selenium, selenide or organic selenium. In this paper, we aim to exclusively bio-reduce selenate to selenite in an open high-rate bioreactor. This conversion could be part of a two-stage process in which the selenite is subsequently reduced by chemical means under optimal conditions to produce a biomass-free selenium product. In the process, yield and reduction rate of biological selenate to selenite should be maximized, while formation of elemental selenium, selenide and organic selenium compounds should be avoided. Fed-batch experiments with a liquid volume of 0.25-0.75 L at different temperatures 20-30-40-50 °C, pH settings 6-7-8-9, initial biomass concentration of 1 or 5 g wet weight granular Eerbeek sludge and various lactic acid concentrations were performed to determine their effect on the biological conversion of selenate to selenite. Furthermore, the effect on selenite losses by further biological reduction or, if present, chemical reduction was investigated as well. Optimal selenate reduction to selenite was found at 30 °C and pH 6 or 7 or 8 with 25 mM selenate and 13.75 mM lactic acid in the influent, with a selenite yield of 79-95%. In all the other conditions, less selenate was reduced to selenite. Also a 5 times higher electron donor concentration resulted in less selenite production, with only 22% of the selenate converted to selenite. The high yield and the high biological reduction rate of at least 741 mg Se/g initial VSS/day detected in the 1 g initial biomass experiment implicate that biological selenate conversion to selenite is a feasible process.

Energy-state dependent responses of Anopheles gambiae (Diptera: Culicidae) to simulated bednet-protected hosts.

In nature, Anopheles gambiae mosquitoes are found at various energy levels and such females must choose between seeking somatic energy from sugar sources and obtaining both somatic and gametic energy from blood hosts. We used a straight-tube olfactometer containing a simulated unobtainable blood host (human foot smell protected by a net) as well as a sugar source (honey odor). We assessed female probing rate and residence time at the net as a function of energy state (0, 24, 48, 72-h starved). In our trials, 0-h starved females showed low response to human odor, low probing rate, and residence time at the human odor site. By contrast, both 48 and 72-h individuals showed high response to foot odor, longer residence time, and higher probing rates. Seventy-two-h females also flew towards the honey source less often than other groups. Our findings suggest that managing sugar sources might be a viable strategy for influencing mosquito biting behavior.

Mining and validation of pyrosequenced simple sequence repeats (SSRs) from American cranberry (Vaccinium macrocarpon Ait.).

The American cranberry (Vaccinium macrocarpon Ait.) is a major commercial fruit crop in North America, but limited genetic resources have been developed for the species. Furthermore, the paucity of codominant DNA markers has hampered the advance of genetic research in cranberry and the Ericaceae family in general. Therefore, we used Roche 454 sequencing technology to perform low-coverage whole genome shotgun sequencing of the cranberry cultivar 'HyRed'. After de novo assembly, the obtained sequence covered 266.3 Mb of the estimated 540-590 Mb in cranberry genome. A total of 107,244 SSR loci were detected with an overall density across the genome of 403 SSR/Mb. The AG repeat was the most frequent motif in cranberry accounting for 35% of all SSRs and together with AAG and AAAT accounted for 46% of all loci discovered. To validate the SSR loci, we designed 96 primer-pairs using contig sequence data containing perfect SSR repeats, and studied the genetic diversity of 25 cranberry genotypes. We identified 48 polymorphic SSR loci with 2-15 alleles per locus for a total of 323 alleles in the 25 cranberry genotypes. Genetic clustering by principal coordinates and genetic structure analyzes confirmed the heterogeneous nature of cranberries. The parentage composition of several hybrid cultivars was evident from the structure analyzes. Whole genome shotgun 454 sequencing was a cost-effective and efficient way to identify numerous SSR repeats in the cranberry sequence for marker development.

Assessment of a computerised decision support system for allergic rhino-conjunctivitis counselling in German pharmacy.

OBJECTIVE: In a prospective two-phase intervention study we evaluated a newly developed computerised pharmacy decision support system (PDSS) for the counselling of patients with allergic rhinitis and conjunctivitis. METHOD: Community pharmacists were invited to counsel a virtual patient in a first interview. Immediately thereafter, we trained pharmacists to operate the PDSS, which they applied in a second patient interview immediately following the training. We assessed the completeness of mandatory questions asked as defined by national guidelines. RESULTS: Participating pharmacists (n = 50, 78% female, mean age 34 [IQR: 27-40] years, 8 [2-14.5] years of practical experience, 32% specialised in community pharmacy) asked considerably more mandatory questions to confirm appropriateness of self-medication with 7 (5.25-9; 78%) from 9 questions compared to 2 (1-3; 22%) without PDSS (median; P < 0.001). In particular, using the PDSS more than doubled mandatory questions (9/12, 6.25-10; 75% vs. 4/12, 3-5; 33%; P < 0.001) relevant for appropriate drug selection. CONCLUSION: Pharmacists omitted many questions mandatory to assess whether self-medication is appropriate. Using the newly developed PDSS more than doubled the number of mandatory questions asked. The results suggest that the PDSS is ready for evaluation of its impact in real patients.

Misspellings in drug information system queries: characteristics of drug name spelling errors and strategies for their prevention.

INTRODUCTION: Efficient search for and finding drugs is essential for electronic drug information systems which, for their part, are prerequisites for computerized physician order entry systems and clinical decision support with the potential to prevent medication errors. Search failures would be critical: they may delay or even prohibit prescription processes or timely retrieval of vital drug information. We analyzed spelling-correction and error characteristics in drug searches and the suitability of auto-completion as prevention strategy. METHODS: A blank entry field was presented to the user for unbiased queries in a web-based drug information system containing >105,000 brand names and active ingredients accessible from all 5500 computers of the Heidelberg University Hospital. The system was equipped with an error-tolerant search. Misspelled but found drug names confirmed by users were aligned by dynamic programming algorithms, opposing misspelled and correct names letter by letter. We analyzed the ratios of correctly and incorrectly spelled but found drugs, frequencies of characters, and their position in misspelled search words. RESULTS: Without error-tolerant search, no results were found in 17.5% of all queries. Users confirmed 31% of all results found with phonetic error-correction support. Sixteen percent of all spelling errors were letters in close proximity to the correct letter on keyboards. On average, 7% of the initial letters in misspelled words contained errors. CONCLUSION: Drug information systems should be equipped with error-tolerant algorithms to reduce search failures. Drug initial letters are also error-prone, thus auto-completion is not a sufficient error-prevention strategy and needs additional support by error-tolerant algorithms.

Multidisciplinary pain management based on a computerized clinical decision support system in cancer pain patients.

A prospective controlled intervention cohort study in cancer pain patients (n=50 per group) admitted to radiation oncology wards (62 beds, 3 wards) was conducted in a 1621-bed university hospital. We investigated the effect of an intervention consisting of daily pain assessment using the numeric visual analog scale (NVAS) and pain therapy counseling to clinicians based on a computerized clinical decision support system (CDSS) to correct deviations from pain therapy guidelines. Effects on guideline adherence (primary outcome), pain relief (NVAS) at rest and during physical activity (both groups: cross-sectional assessment on day 5; intervention group: every day assessment), co-analgesic prescription, and acceptance rates of recommendations (secondary outcomes) were assessed. The number of patients with at least one deviation from guidelines at discharge was decreased by the intervention from 37 (74%) in controls to 7 (14%, p<0.001). In the intervention group, pain (NVAS) decreased during hospital stay at rest from 3.0 (Delta(0.5) (Q(75%)-Q(25%))=3.0) on admission to 1.5 (Delta(0.5)=1.0) at discharge (p<0.01) and during physical activity from 7.0 (Delta(0.5)=4.0) on admission to 2.5 (Delta(0.5)=3.8) at discharge (p<0.001). At discharge, the number of patients treated with co-analgesics increased from 23 (46%) in controls to 33 (66%) in the intervention group (p=0.04). From 279 recommendations issued in the intervention 85% were fully accepted by the physicians. Deviations from well-established guidelines are frequent in pain therapy. A multidisciplinary pain management increased adherence to pain management guidelines.

Substantial reduction of inappropriate tablet splitting with computerised decision support: a prospective intervention study assessing potential benefit and harm.

BACKGROUND: Currently ambulatory patients break one in four tablets before ingestion. Roughly 10% of them are not suitable for splitting because they lack score lines or because enteric or modified release coating is destroyed impairing safety and effectiveness of the medication. We assessed impact and safety of computerised decision support on the inappropriate prescription of split tablets. METHODS: We performed a prospective intervention study in a 1680-bed university hospital. Over a 15-week period we evaluated all electronically composed medication regimens and determined the fraction of tablets and capsules that demanded inappropriate splitting. In a subsequent intervention phase of 15 weeks duration for 10553 oral drugs divisibility characteristics were indicated in the system. In addition, an alert was generated and displayed during the prescription process whenever the entered dosage regimen demanded inappropriate splitting (splitting of capsules, unscored tablets, or scored tablets unsuitable for the intended fragmentation). RESULTS: During the baseline period 12.5% of all drugs required splitting and 2.7% of all drugs (257/9545) required inappropriate splitting. During the intervention period the frequency of inappropriate splitting was significantly reduced (1.4% of all drugs (146/10486); p = 0.0008). In response to half of the alerts (69/136) physicians adjusted the medication regimen. In the other half (67/136) no corrections were made although a switch to more suitable drugs (scored tablets, tablets with lower strength, liquid formulation) was possible in 82% (55/67). CONCLUSION: This study revealed that computerised decision support can immediately reduce the frequency of inappropriate splitting without introducing new safety hazards.

A consensus linkage map identifies genomic regions controlling fruit maturity and beta-carotene-associated flesh color in melon (Cucumis melo L.).

The nutritional value and yield potential of US Western Shipping melon (USWS; Cucumis melo L.) could be improved through the introgression of genes for early fruit maturity (FM) and the enhancement of the quantity of beta-carotene (QbetaC) in fruit mesocarp (i.e., flesh color). Therefore, a set of 116 F(3) families derived from the monoecious, early FM Chinese line 'Q 3-2-2' (no beta-carotene, white mesocarp) and the andromonoecious, late FM USWS line 'Top Mark' (possessing beta-carotene, orange mesocarp) were examined during 2 years in Wisconsin, USA to identify quantitative trait loci (QTL) associated with FM and QbetaC. A 171-point F(2-3) based map was constructed and used for QTL analysis. Three QTL associated with QbetaC were detected, which explained a significant portion of the observed phenotypic variation (flesh color; R (2) = 4.0-50.0%). The map position of one QTL (beta-carM.E.9.1) was uniformly aligned with one carotenoid-related gene (Orange gene), suggesting its likely role in QbetaC in this melon population and putative relationship with the melon white flesh (wf) gene. Two major (FM.6.1 and FM.11.1; R (2) >or= 20%) and one minor QTL (FM.2.1; R (2) = 8%) were found to be associated with FM. This map was then merged with a previous recombinant inbred line (RIL)-based map used to identify seven QTL associated with QbetaC in melon fruit. This consensus map [300 molecular markers (187 co-dominant melon and 14 interspecific; 10 LG)] provides a framework for the further dissection and cloning of published QTL, which will consequently lead to more effective trait introgression in melon.

Mapping of genetic loci that regulate quantity of beta-carotene in fruit of US Western Shipping melon (Cucumis melo L.).

Melon (Cucumis melo L.) is highly nutritious vegetable species and an important source of beta-carotene (Vitamin A), which is an important nutrient in the human diet. A previously developed set of 81 recombinant inbred lines (RIL) derived from Group Cantalupensis US Western Shipper market type germplasm was examined in two locations [Wisconsin (WI) and California (CA), USA] over 2 years to identify quantitative trait loci (QTL) associated with quantity of beta-carotene (QbetaC) in mature fruit. A moderately saturated 256-point RIL-based map [104 SSR, 7 CAPS, 4 SNP in putative carotenoid candidate genes, 140 dominant markers and one morphological trait (a) spanning 12 linkage groups (LG)] was used for QbetaC-QTL analysis. Eight QTL were detected in this evaluation that were distributed across four LG that explained a significant portion of the associated phenotypic variation for QbetaC (R (2) = 8 to 31.0%). Broad sense heritabilities for QbetaC obtained from RIL grown in WI. and CA were 0.56 and 0.68, respectively, and 0.62 over combined locations. The consistence of QbetaC in high/low RIL within location across years was confirmed in experiments conducted over 2 years. QTL map positions were not uniformly associated with putative carotenoid genes, although one QTL (beta-car6.1) interval was located 10 cM from a beta-carotene hydroxylase gene. These results suggest that accumulation of beta-carotene in melon is under complex genetic control. This study provides the initial step for defining the genetic control of QbetaC in melon leading to the development of varieties with enhanced beta-carotene content.

Characteristics of effective electronic mail messages distributed to healthcare professionals in a hospital setting.

Electronic mailing systems (e-mail) are an important means to disseminate information within electronic networks. However, in large business communities including the hectic environment of hospitals it may be difficult to induce account holders to read the e-mail. In two mailings disseminated in a large university hospital we evaluated the impact of e-mail layout (three e-mail text versions, two e-mails with graphics) on the willingness of its approximately 6500 recipients to seek additional electronic information and open an integrated link. Overall access rates after 90 days were 21.1 and 23.5% with more than 70% of the respondents opening the link within 3 days. Differences between different layouts were large and artwork text, HTML text, animated GIF, and static image prompted 1.2, 1.7, 1.8, and 2.3 times more often access than the courier plain text message (p

Carotenoid biosynthesis structural genes in carrot (Daucus carota): isolation, sequence-characterization, single nucleotide polymorphism (SNP) markers and genome mapping.

Carotenoid pigments are important components of the human diet and carrots are the main dietary sources of the vitamin A precursors alpha- and beta-carotene. Carotenoids play essential biological roles in plants and the genes coding for the carotenoid pathway enzymes are evolutionarily conserved, but little information exists about these genes for carrot. In this study, we utilized published carrot sequences as well as heterologous PCR approaches with primers derived from sequence information of other plant species to isolate 24 putative genes coding for carotenoid biosynthesis enzymes in carrot. Twenty-two of these genes were placed on the carrot genetic linkage map developed from a cross between orange-rooted and white-rooted carrot. The carotenoid genes were distributed in eight of the nine linkage groups in the carrot genome recommending their use for merging maps. Two genes co-localized with a genomic region spanning one of the most significant quantitative trait loci (QTL) for carotenoid accumulation. Carotenoid biosynthesis cDNAs linked to root color mutations and to QTL for carotenoid accumulation may suggest a functional role for them as candidate genes. RACE PCR and reverse transcriptase PCR were used to amplify the full-length transcript for twenty expressed carotenoid biosynthesis genes and sequences were submitted to GenBank. The cloning and sequence information of these genes is useful for PCR-based expression studies and may point toward transgenic approaches to manipulate carotenoid content in carrot.

Single seed Raman measurements allow taxonomical discrimination of Apiaceae accessions collected in gene banks.

NIR-FT-Raman spectroscopy was applied for a nondestructive analysis of single seeds (fruit mericarps) of 36 accessions belonging to various species of the Apiaceae family. Main seed components such as fatty acids, polysaccharides, proteins, and lignin were identified based on the obtained Raman spectra. Variation at the species and genus level was related to differences observed between spectra. The application of cluster analysis discriminated among most of the species evaluated and grouped them according to their taxonomical classification. The spectroscopically analyzed seeds germinated and developed into normal seedlings to demonstrate the additional advantage that Raman spectroscopy is nondestructive and can be applied to living seed without harm. These results indicate that Raman spectroscopy is a valuable tool for the rational evaluation and management of genetic resources in ex situ seed collections by providing useful information for taxonomical validation of the accessions.

Demonstration of linkage and development of the first low-density genetic map of garlic, based on AFLP markers.

Garlic (Allium sativum L.) is a long-cultivated, clonally propagated diploid plant (2n=2x=16). With routine seed production now underway, we used populations (MP1 and MP2) generated by self-pollination of unrelated plants to generate two low-density genetic maps of garlic, consisting of amplified fragment length polymorphism (AFLP) and gene-specific markers. We did not observe any two plants with identical marker patterns in either population, indicating that they were the result of amphimixis rather than apomixis. This is an important finding, since several Alliums are facultative apomicts. A total of 360 markers segregated in MP1 (12.8 AFLP markers per primer combination) and 321 markers segregated in MP2 (13.9 per primer combination) to indicate a fairly high level of genetic heterozygosity in the garlic nuclear genome. Of these markers, 15.3% in MP1 and 24.3% in MP2 had segregation ratios distorted from the expected 3:1. Interestingly, 94.7% of those distorted segregations fit a 15:1 segregation ratio for duplicated loci, suggesting extensive levels of duplication in the garlic genome and supporting similar observations for onion. The genetic map for the MP1 family with 216 markers spanned 1,166 cM of the garlic genome (5.4 cM average), while 143 markers of MP2 spanned 862 cM (6.0 cM average). Gene-specific markers for alliinase, chitinase, sucrose 1-fructosyltransferase (SST-1), and chalcone synthase (CHS) were mapped, demonstrating the immediate utility of the garlic genetic map. These two garlic families had relatively few segregating AFLP markers in common, which supports their relatively distant relationship based on diversity analysis. Of those markers that were conserved, linkages were also conserved.

Lycopene and beta-carotene are bioavailable from lycopene 'red' carrots in humans.

OBJECTIVE: The purpose of this study was to determine if lycopene and beta-carotene are bioavailable from lycopene red carrots and if lycopene absorption is affected by carrot fiber. DESIGN: Two crossover studies in humans attempted to compare the relative bioavailability of lycopene and beta-carotene from tomato paste to a genetically selected lycopene red carrot during chronic feeding. Each study contained three treatment groups. The vehicle of administration was muffins. INTERVENTION AND METHODS: Study 1 (n=9) used white carrots (0 mg lycopene/day), red carrots (5 mg/day), and tomato paste (20 mg/day). Study 2 (n=10) used red carrots (2.6 mg/day), tomato paste (5 mg/day), and tomato paste plus white carrots (5 mg/day). Each intervention lasted 11 days with a 10-day washout period between treatments. Serum lycopene and beta-carotene were measured by HPLC. RESULTS: Statistical analysis indicated a significant effect of muffin type in study 1 (P<0.001), and a significant treatment by sequence interaction in study 2 (P=0.04). The response to increasing amounts of lycopene is linear at the levels fed in these studies (r=0.94). The data suggest that maintenance of serum lycopene concentrations at 0.3 micromol/l occurs at about 2 mg/day of lycopene from mixed dietary sources and a serum plateau occurs at >/=20 mg/day. CONCLUSIONS: These results show that lycopene and beta-carotene are bioavailable from red carrots and lycopene absorption seems to be affected by carrot fiber. Making inferences from both studies, the lycopene in the red carrot is about 44% as bioavailable as that from tomato paste. Red carrots provide an alternative to tomato paste as a good dietary source of lycopene and also provide bioavailable beta-carotene.

QTL analyses reveal clustered loci for accumulation of major provitamin A carotenes and lycopene in carrot roots.

QTLs associated with products of the carotenoid pathway, including lycopene and the provitamin A carotenes alpha- and beta-carotene, were investigated in two unrelated F(2) carrot populations, derived from crosses between orange cultivated B493 and white wild QAL (Population 1), and orange cultivated Brasilia and dark-orange cultivated HCM (Population 2). The mapping populations of 160 and 180 individuals, respectively, were analyzed with single-marker and interval-mapping statistical approaches, using coupling linkage maps for each parent. Single markers were selected for further analysis based on the Wilcoxon sum-rank non-parametric test. Interval mapping performed with Population 2 detected four, eight, three, one and five putative QTLs associated with accumulation of xi-carotene, alpha-carotene, beta-carotene, lycopene and phytoene, respectively. Among these, the major QTLs explained 13.0%, 10.2%, 13.0%, 7.2% and 10.2% of total phenotypic variation. In Population 1 single-marker analysis identified loci explaining up to 13.8%, 6.8%, 19.3%, 5.7%, and 17.5%, respectively, of total phenotypic variation for these same carotenoids. Overall analysis demonstrated clustering of these QTLs associated with the carotenoid pathway: the AFLP loci AACCAT178-Q and AAGCAG233-Q, on linkage group 5, explained 17.8%, 22.8% and 23.5% of total phenotypic variation for zeta-carotene, phytoene and beta-carotene in Population 1. Two major clusters of QTLs, with LOD scores greater than 1.8, mapped to intervals no larger than 2 cM for zeta-carotene, beta-carotene, alpha-carotene and lycopene on linkage group 3, and for zeta-carotene and phytoene on linkage group 9, and these explained 3.7% to 13.0% of variation for each carotenoid product. Thus, these results suggest that clustering of related pathway loci is favored during evolution, since closely linked "pathway mates" are not easily separated by recombination.

Improved phytonutrient content through plant genetic improvement.

During the twentieth century, plant breeding and genetics improved the nutritive value of horticultural and agronomic crops, particularly of macronutrients and fiber. Current research focuses more on micronutrients. Successful development of phytonutrient-enriched crop plants will be bolstered by interdisciplinary collaborative research, analytical and biotechnology advances, and public education. Although the melding of plant and nutrition research holds great promise, the genetic enhancement of crop plants for improved phytonutrient content will be challenging. This paper reviews the knowledge base on which genetic enhancement may be based, identifies gaps in scientific knowledge and technical capacities, and suggests a role for the federal government in research.

Regeneration of garlic callus as affected by clonal variation, plant growth regulators and culture conditions over time.

A long-term regeneration system for garlic (Allium sativum L.) clones of diverse origin was developed. Callus was initiated on a modified Gamborg's B-5 medium supplemented with 4.5 µM 2,4-D and maintained on the same basal medium with 4.7 µM picloram+0.49 µM 2iP. Regeneration potential of callus after 5, 12 and 16 months on maintenance medium was measured using several plant growth regulator treatments. The 1.4 µM picloram+13.3 µM BA treatment stimulated the highest rate of shoot production. Regeneration rate decreased as callus age increased, but healthy plantlets from callus cultures up to 16-months-old were produced for all clones. Regeneration of long-term garlic callus cultures could be useful for clonal propagation and transformation.