856 resultados para anatomical and physiological traits


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Movement of tephritid flies underpins their survival, reproduction, and ability to establish in new areas and is thus of importance when designing effective management strategies. Much of the knowledge currently available on tephritid movement throughout landscapes comes from the use of direct or indirect methods that rely on the trapping of individuals. Here, we review published experimental designs and methods from mark-release-recapture (MRR) studies, as well as other methods, that have been used to estimate movement of the four major tephritid pest genera (Bactrocera, Ceratitis, Anastrepha, and Rhagoletis). In doing so, we aim to illustrate the theoretical and practical considerations needed to study tephritid movement. MRR studies make use of traps to directly estimate the distance that tephritid species can move within a generation and to evaluate the ecological and physiological factors that influence dispersal patterns. MRR studies, however, require careful planning to ensure that the results obtained are not biased by the methods employed, including marking methods, trap properties, trap spacing, and spatial extent of the trapping array. Despite these obstacles, MRR remains a powerful tool for determining tephritid movement, with data particularly required for understudied species that affect developing countries. To ensure that future MRR studies are successful, we suggest that site selection be carefully considered and sufficient resources be allocated to achieve optimal spacing and placement of traps in line with the stated aims of each study. An alternative to MRR is to make use of indirect methods for determining movement, or more correctly, gene flow, which have become widely available with the development of molecular tools. Key to these methods is the trapping and sequencing of a suitable number of individuals to represent the genetic diversity of the sampled population and investigate population structuring using nuclear genomic markers or non-recombinant mitochondrial DNA markers. Microsatellites are currently the preferred marker for detecting recent population displacement and provide genetic information that may be used in assignment tests for the direct determination of contemporary movement. Neither MRR nor molecular methods, however, are able to monitor fine-scale movements of individual flies. Recent developments in the miniaturization of electronics offer the tantalising possibility to track individual movements of insects using harmonic radar. Computer vision and radio frequency identification tags may also permit the tracking of fine-scale movements by tephritid flies by automated resampling, although these methods come with the same problems as traditional traps used in MRR studies. Although all methods described in this chapter have limitations, a better understanding of tephritid movement far outweighs the drawbacks of the individual methods because of the need for this information to manage tephritid populations.

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The present research focused on motivational and personality traits measuring individual differences in the experience of negative affect, in reactivity to negative events, and in the tendency to avoid threats. In this thesis, such traits (i.e., neuroticism and dispositional avoidance motivation) are jointly referred to as trait avoidance motivation. The seven studies presented here examined the moderators of such traits in predicting risk judgments, negatively biased processing, and adjustment. Given that trait avoidance motivation encompasses reactivity to negative events and tendency to avoid threats, it can be considered surprising that this trait does not seem to be related to risk judgments and that it seems to be inconsistently related to negatively biased information processing. Previous work thus suggests that some variable(s) moderate these relations. Furthermore, recent research has suggested that despite the close connection between trait avoidance motivation and (mal)adjustment, measures of cognitive performance may moderate this connection. However, it is unclear whether this moderation is due to different response processes between individuals with different cognitive tendencies or abilities, or to the genuinely buffering effect of high cognitive ability against the negative consequences of high trait avoidance motivation. Studies 1-3 showed that there is a modest direct relation between trait avoidance motivation and risk judgments, but studies 2-3 demonstrated that state motivation moderates this relation. In particular, individuals in an avoidance state made high risk judgments regardless of their level of trait avoidance motivation. This result explained the disparity between the theoretical conceptualization of avoidance motivation and the results of previous studies suggesting that the relation between trait avoidance motivation and risk judgments is weak or nonexistent. Studies 5-6 examined threat identification tendency as a moderator for the relationship between trait avoidance motivation and negatively biased processing. However, no evidence for such moderation was found. Furthermore, in line with previous work, the results of studies 5-6 suggested that trait avoidance motivation is inconsistently related to negatively biased processing, implying that theories concerning traits and information processing may need refining. Study 7 examined cognitive ability as a moderator for the relation between trait avoidance motivation and adjustment, and demonstrated that cognitive ability moderates the relation between trait avoidance motivation and indicators of both self-reported and objectively measured adjustment. Thus, the results of Study 7 supported the buffer explanation for the moderating influence of cognitive performance. To summarize, the results showed that it is possible to find factors that consistently moderate the relations between traits and important outcomes (e.g. adjustment). Identifying such factors and studying their interplay with traits is one of the most important goals of current personality research. The present thesis contributed to this line of work in relation to trait avoidance motivation.

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Tephritid fruit flies (Diptera: Tephritidae) are considered by far the most important group of horticultural pests worldwide. Female fruit flies lay eggs directly into ripening fruit, where the maggots feed causing fruit loss. Each and every continent is plagued by a number of fruit fly pests, both indigenous as well as invasive ones, causing tremendous economic losses. In addition to the direct losses through damage, they can negatively impact commodity trade through restrictions to market access. The quarantine and regulatory controls put in place to manage them are expensive, while the on-farm control costs and loss of crop affect the general well-being of growers. These constraints can have huge implications on loss in revenues and limitations to developing fruit and vegetable-based agroindustries in developing, emergent and developed nations. Because fruit flies are a global problem, the study of their biology and management requires significant international attention to overcome the hurdles they pose. The Joint Food and Agriculture Organisation / International Atomic Energy Agency (FAO/IAEA) Programme on Nuclear Techniques in Food and Agriculture has been on the foreground in assisting Member States in developing and validating environment-friendly fruit fly suppression systems to support viable fresh fruit and vegetable production and export industries. Such international attention has resulted in the successful development and validation of a Sterile Insect Technique (SIT) package for the Mediterranean fruit fly. Although demands for R&D support with respect to Mediterranean fruit fly are diminishing due to successful integration of this package into sustainable control programmes against this pest in many countries, there were increasing demands from Member States in Africa, Asia and Latin America, to address other major fruit fly pests and a related, but sometimes neglected issue of tephritid species complexes of economic importance. Any research, whether it is basic or applied, requires a taxonomic framework that provides reliable and universally recognized entities and names. Among the currently recognized major fruit fly pests, there are groups of species whose morphology is very similar or identical, but biologically they are distinct species. As such, some insect populations that are grouped taxonomically within the same pest species, display different biological and genetic traits and show reproductive isolation which suggest that they are different species. On the other hand, different species may have been taxonomically described, but there may be doubt as to whether they actually represent distinct biological species or merely geographical variants of the same species. This uncertain taxonomic status has practical implications on the effective development and use of the SIT against such complexes, particularly at the time of determining which species to mass-rear, and significantly affects international movement of fruit and vegetables through the establishment of trade barriers to important agricultural commodities which are hosts to these pest tephritid species...

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Two laboratory experiments were carried out to quantify the mortality and physiological responses of juvenile blue swimmer crabs (Portunus pelagicus) after simulated gillnet entanglement, air exposure, disentanglement, and discarding. In both experiments, all but control blue swimmer crabs were entangled in 1-m(2) gillnet panels for 1 h, exposed to air for 2 min, subjected to various treatments of disentanglement ranging between the forceful removal of none, one, two, and four appendages, then "discarded" into individual experimental tanks and monitored for 10 d. In Experiment 1, mortalities were associated with the number of appendages removed and the occurrence of unsealed wounds. In Experiment 2, live blue swimmer crabs were sampled for blood at 2 min and 6, 24, and 72 h post-discarding to test for the effects of disentanglement and appendage removal on total haemocyte counts, clotting times, protein levels (by refractive index), and blood ion concentrations. Compared with blue swimmer crabs that had sealed or no wounds, those with unsealed wounds had lower total haemocyte counts, protein, and calcium concentrations and increased clotting ties and magnesium and sodium levels. Induced autotomy, as opposed to the arbitrary, forceful removal of a appendages has the potential to minimize the mortality and stress of discarded, juvenile blue swimmer crabs.

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Purpose Transient changes in corneal topography associated with soft and conventional or reverse geometry rigid contact lens wear have been well documented; however, only a few studies have examined the influence of scleral contact lens wear upon the cornea. Therefore, in this study, we examined the influence of modern miniscleral contact lenses, which land entirely on the sclera and overlying tissues, upon anterior corneal curvature and optics. Methods Anterior corneal topography and elevation data were acquired using Scheimpflug imaging (Pentacam HR, Oculus) immediately prior to and following 8 hours of miniscleral contact lens wear in 15 young healthy adults (mean age 22 ± 3 years, 8 East Asian, 7 Caucasian) with normal corneae. Corneal diurnal variations were accounted for using data collected on a dedicated measurement day without contact lens wear. Corneal clearance was quantified using an optical coherence tomographer (RS-3000, Nidek) following lens insertion and after 8 hours of lens wear. Results Although corneal clearance was maintained throughout the 8 hour lens wear period, significant corneal flattening (up to 0.08 ± 0.04 mm) was observed, primarily in the superior mid-peripheral cornea, which resulted in a slight increase in against-the-rule corneal astigmatism (mean +0.02/-0.15 x 94 for an 8 mm diameter). Higher order aberration terms of horizontal coma, vertical coma and spherical aberration all underwent significant changes for an 8 mm corneal diameter (p ≤ 0.01), which typically resulted in a decrease in RMS error values (mean change in total higher order RMS -0.035 ± 0.046 µm for an 8 mm diameter). There was no association between the magnitude of change in central or mid-peripheral corneal clearance during lens wear and the observed changes in corneal curvature (p > 0.05). However, Asian participants displayed a significantly greater reduction in corneal clearance (p = 0.04) and greater superior-nasal corneal flattening compared to Caucasians (p = 0.048). Conclusions Miniscleral contact lenses that vault the cornea induce significant changes in anterior corneal surface topography and higher order aberrations following 8 hours of lens wear. The region of greatest corneal flattening was observed in the superior-nasal mid-periphery, more so in Asian participants. Practitioners should be aware that corneal measurements obtained following miniscleral lens removal may mask underlying corneal steepening.

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The genus Asparagus includes at least six invasive species in Australia. Asparagus aethiopicus and A. africanus are invasive in subtropical Australia, and a third species, A. virgatus is naturalized and demonstrates localized spread in south east Queensland. To better understand how the attributes of these species contribute to their invasiveness, we compared fruit and seed traits, germination, seedling emergence, seed survival, and time-to-maturity. We further investigated dispersal ecology of A. africanus, examining the diet of a local frugivore, the figbird (Sphecotheres viridis) and the effect of gut passage on seedling emergence. Overall, A. aethiopicus was superior in germination and emergence, with the highest mean germination (98.8%) and emergence (94.5%) under optimal conditions and higher emergence (mean of 73.3%) across all treatments. In contrast, A. africanus had the lowest germination under optimal conditions (71.7%) and low mean seedling emergence (49.5%), but had fruits with the highest relative yield (ratio of dry pulp to fruit fresh weight) that were favored by a local frugivore. Figbirds consumed large numbers of A. africanus fruits (~30% of all non-Ficus fruits), and seedling germination was not significantly affected by gut passage compared to unprocessed fruits. Asparagus virgatus germinated poorly under cool, light conditions (1.4%) despite a high optimum mean (95.0%) and had low mean performance across emergence treatments (36.3%). The species also had fruits with a low pulp return for frugivores. For all species, seed survival declined rapidly in the first 12 mo and fell to < 3.2% viability at 36 mo. On the basis of the traits considered, A. virgatus is unlikely to have the invasive potential of its congeners. Uniformly short seed survival times suggest that weed managers do not have to contend with a substantial persistent soil-stored seed bank, but frugivore-mediated dispersal beyond existing infestations will present a considerable management challenge.

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Characterisation and investigation of a number of key wood properties, critical for further modelling work, has been achieved. The key results were: • Morphological characterisation, in terms of fibre cell wall thickness and porosity, was completed. A clear difference in fibre porosity, size, wall thickness and orientation was evident between species. Results were consistent with published data for other species. • Viscoelastic properties of wood were shown to differ greatly between species and in the radial and tangential directions, largely due to anatomical and chemical variations. Consistent with published data, the radial direction shows higher stiffness, internal friction and glass transition temperature than the tangential directions. The loss of stiffness over the measured temperature range was greater in the tangential direction than the radial direction. Due to time dependant molecular relaxation, the storage modulus and glass transition temperature decreased with decreasing test frequency, approaching an asymptotic limit. Thus the viscoelastic properties measured at lower frequencies are more representative of static material. • Dynamic interactions between relative humidity, moisture content and shrinkage of four Australian hardwood timbers can be accurately monitored on micro-samples using a specialised experimental device developed by AgroParisTech – ENGREF. The device generated shrinkage data that varied between species but were consistent (repeatable) within a species. Collapse shrinkage was clearly evident with this method for Eucalyptus obliqua, but not with other species, consistent with industrial seasoning experience. To characterise the wood-water relations of this species, free of collapse, thinner sample sections (in the R-T plane) should be used.

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Cardiovascular diseases (CVD) are a major cause of death and disability in Western countries and a growing health problem in the developing world. The genetic component of both coronary heart disease (CHD) and ischemic stroke events has been established in twin studies, and the traits predisposing to CVD, such as hypertension, dyslipidemias, obesity, diabetes, and smoking behavior, are all partly hereditary. Better understanding of the pathophysiology of CVD-related traits could help to target disease prevention and clinical treatment to individuals at an especially high disease risk and provide novel pharmaceutical interventions. This thesis aimed to clarify the genetic background of CVD at a population level using large Nordic population cohorts and a candidate gene approach. The first study concentrated on the allelic diversity of the thrombomodulin (THBD) gene in two Finnish cohorts, FINRISK-92 and FINRISK-97. The results from this study implied that THBD variants do not substantially contribute to CVD risk. In the second study, three other candidate genes were added to the analyses. The study investigated the epistatic effects of coagulation factor V (F5), intercellular adhesion molecule -1 (ICAM1), protein C (PROC), and THBD in the same FINRISK cohorts. The results were encouraging; we were able to identify several single SNPs and SNP combinations associating with CVD and mortality. Interestingly, THBD variants appeared in the associating SNP combinations despite the negative results from Study I, suggesting that THBD contributes to CVD through gene-gene interactions. In the third study, upstream transcription factor -1 (USF1) was analyzed in a cohort of Swedish men. USF1 was associated with metabolic syndrome, characterized by accumulation of different CVD risk factors. A putative protective and a putative risk variant were identified. A direct association with CVD was not observed. The longitudinal nature of the study also clarified the effect of USF1 variants on CVD risk factors followed in four examinations throughout adulthood. The three studies provided valuable information on the study of complex traits, highlighting the use of large study samples, the importance of replication, and the full coverage of the major allelic variants of the target genes to assure reliable findings. Although the genetic basis of coronary heart disease and ischemic stroke remains unknown, single genetic findings may facilitate the recognition of high-risk subgroups.

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The objectives of this study were to quantify the components of genetic variance and the genetic effects, and to examine the genetic relationship of inbred lines extracted from various shrunken2(sh2) breeding populations. Ten diverse inbred lines developed from genetic background, were crossed in half diallel. Parents and their F1 hybrids were evaluated at three environments. The parents were genotyped using 20 polymorphic simple sequence repeats (SSR). Agronomic and quality traits were analysed by a mixed linear model according to additive-dominance genetic model. Genetic effects were estimated using an adjusted unbiased prediction method. Additive variance was more important than dominance variance in the expression of traits related to ear aspects (husk ratio and percentage of ear filled) and eating quality (flavour and total soluble solids). For agronomic traits, however, dominance variance was more important than additive variance. The additive genetic correlation between flavour and tenderness was strong (r = 0.84, P <0.01). Flavour, tenderness and kernel colour additive genetic effects were not correlated with yield related traits. Genetic distance (GD), estimated from SSR profiles on the basis of Jaccard's similarity coefficient varied from 0.10 to 0.77 with an average of 0.56. Cluster analysis classified parents according to their pedigree relationships. In most studied traits, F1 performance was not associated with GD.

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The effect of nonresponse on health and lifestyle measures has received extensive study, showing at most relatively modest effects. Nonresponse bias with respect to personality has been less thoroughly investigated. The present study uses data from responding individuals as a proxy for the missing data of their nonresponding family members to examine the presence of nonresponse bias for personality traits and disorders as well as health and lifestyle traits. We looked at the Big Five personality traits, borderline personality disorder (BPD) features, attention-deficit/hyperactivity disorder, Anger, and several measures of health (Body Mass Index, migraine) and lifestyle (smoking, alcohol use). In general, outcomes tend to be slightly more favorable for individuals from highly cooperative families compared to individuals from less cooperative families. The only significant difference was found for BPD features (p = .001). However, the absolute difference in mean scores is very small, less than 1 point for a scale ranging from 0 to 72. In conclusion, survey data on personality, health and lifestyle are relatively unbiased with respect to nonresponse.

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Performances of Pinus taxa were studied to 10 years of age in two trials in each of Misiones and Entre Rios provinces across the Mesopotamia region of Argentina. Taxa comprised 22 populations from sources in Argentina, Australia, Brazil and Zimbabwe including Pinus elliottii var. elliottii (Pee), Pinus caribaea var. hondurensis (Pch), their four, inter-specific hybrids (F-1, F-2 and backcrosses from F-1 to Pch and to Pee-all as broadly based bulks); other Pee and Pinus taeda (Pt) comprised narrower or unspecified bulks. Variable numbers of taxa were missing at each site. Mean survival across sites at age 10 years ranged 53.2-91.3% averaging 74.2%. Analysis of variance of plot means indicated population effect was statistically significant (p < 0.05) for all or most growth and quality traits at all sites. However, significant differences from the nominated check seedlot at the Entre Rios sites (Pee, Australia) were extremely rare, while quite common at the northern, Misiones sites (check seedlot a Pt population). In the Misiones trials, F-1, F-2 and both backcross hybrids showed better stem straightness than Pee and Pt from Argentina, generally with statistically significant differences (p < 0.05). Pt showed lowest forking scores (desirable). Taxon x environment interaction was statistically significant (p < 0.01) for growth traits only (p > 0.05). However, this interaction contributed an average of only 34.1% of the taxon variance suggesting a lack of practical importance. Taxa most suitable for deployment in the Mesopotamia region, Argentina are suggested.

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Significant interactions have been demonstrated between production factors and postharvest quality of fresh fruit. Accordingly, there is an attendant need for adaptive postharvest actions to modulate preharvest effects. The most significant preharvest effects appear to be mediated through mineral nutrition influences on the physical characteristics of fruit. Examples of specific influencers include fertilisers, water availability, rootstock, and crop load effects on fruit quality attributes such as skin colour, susceptibility to diseases and physiological disorders, and fruit nutritional composition. Also, rainfall before and during harvest can markedly affect fruit susceptibility to skin blemishes, physical damage, and diseases. Knowledge of preharvest-postharvest interactions can help determine the basis for variability in postharvest performance and thereby allow refinement of postharvest practices to minimise quality loss after harvest. This knowledge can be utilised in predictive management systems. Such systems can benefit from characterisation of fruit nutritional status, particularly minerals, several months before and/or at harvest to allow informed decisions on postharvest handling and marketing options. Other examples of proactive management practices include adjusting harvesting and packing systems to account for rainfall effects before and/or during harvest. Improved understanding of preharvest-postharvest interactions is contributing to the delivery of consistently higher quality of fruit to consumers. This paper focuses on the state of knowledge for sub-tropical and tropical fruits, in particular avocado and mango.

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When genome sections of wild Solanum species are bred into the cultivated potato (S. tuberosum L.) to obtain improved potato cultivars, the new cultivars must be evaluated for their beneficial and undesirable traits. Glycoalkaloids present in Solanum species are known for their toxic as well as for beneficial effects on mammals. On the other hand, glycoalkaloids in potato leaves provide natural protection against pests. Due to breeding, glycoalkaloid profile of the plant is affected. In addition, the starch properties in potato tubers can be affected as a result of breeding, because the crystalline properties are determined by the botanical source of the starch. Starch content and composition affect the texture of cooked and processed potatoes. In order to determine glycoalkaloid contents in Solanum species, simultaneous separation of glycoalkaloids and aglycones using reversed-phase high-performance liquid chromatography (HPLC) was developed. Clean-up of foliage samples was improved using a silica-based strong cation exchanger instead of octadecyl phases in solid-phase extraction. Glycoalkaloids alpha-solanine and alpha-chaconine were detected in potato tubers of cvs. Satu and Sini. The total glycoalkaloid concentration of non-peeled and immature tubers was at an acceptable level (under 20 mg/100 g of FW) in the cv. Satu, whereas concentration in cv. Sini was 23 mg/100 g FW. Solanum species (S. tuberosum, S. brevidens, S. acaule, and S. commersonii) and interspecific somatic hybrids (brd + tbr, acl + tbr, cmm + tbr) were analyzed for their glycoalkaloid contents using liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). The concentrations in the tubers of the brd + tbr and acl + tbr hybrids remained under 20 mg/100 g FW. Glycoalkaloid concentration in the foliage of the Solanum species was between 110 mg and 890 mg/100 g FW. However, the concentration in the foliage of S. acaule was as low as 26 mg/100 g FW. The total concentrations of brd + tbr, acl + tbr, and cmm + tbr hybrid foliages were 88 mg, 180 mg, and 685 mg/100 g FW, respectively. Glycoalkaloids of both parental plants as well as new combinations of aglycones and saccharides were detected in somatic hybrids. The hybrids contained mainly spirosolanes, and glycoalkaloid structures having no 5,6-double bond in the aglycone. Based on these results, the glycoalkaloid profiles of the hybrids may represent a safer and more beneficial spectrum of glycoalkaloids than that found in currently cultivated varieties. Starch nanostructure of three different cultivars (Satu, Saturna, and Lady Rosetta), a wild species S. acaule, and interspecific somatic hybrids were examined by wide-angle and small-angle X-ray scattering (WAXS, SAXS). For the first time, the measurements were conducted on fresh potato tuber samples. Crystallinity of starch, average crystallite size, and lamellar distance were determined from the X-ray patterns. No differences in the starch nanostructure between the three different cultivars were detected. However, tuber immaturity was detected by X-ray scattering methods when large numbers of immature and mature samples were measured and the results were compared. The present study shows that no significant changes occurred in the nanostructures of starches resulting from hybridizations of potato cultivars.

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With 6 tables Abstract The objectives of this study were to evaluate the importance of heterosis for agronomic and quality traits in shrunken (sh2) sweet corn, assess the usefulness of combining ability to predict the value of parents and their crosses for further genetic improvement and examine whether genetic divergence can predict heterosis or F1 performance. Ten genetically diverse shrunken (sh2) sweet corn inbred lines were used to generate 45 F1s. F1s and parents were evaluated for agronomic and quality traits across environments. Heterosis was more important for yield-related traits than it was for ear aspects and eating quality. Heterosis for most traits was mostly dependent on dominance genetic effects of parental lines. Parents and F1per se performance were highly correlated with general combining ability effects and mid-parent values, respectively, for most traits. Hybrid performance for flavour and plant height was significantly but weakly related to simple sequence repeat (SSR)-based genetic distance (GD). Phenotypic distance (PD), estimated from phenotypic traits was correlated with heterosis for total soluble solids, ear length and flavour. © 2012 State of Queensland.

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The Queensland strawberry (Fragaria ×ananassa) breeding program in subtropical Australia aims to improve sustainable profitability for the producer. Selection must account for the relative economic importance of each trait and the genetic architecture underlying these traits in the breeding population. Our study used estimates of the influence of a trait on production costs and profitability to develop a profitability index (PI) and an economic weight (i.e., change in PI for a unit change in level of trait) for each trait. The economic weights were then combined with the breeding values for 12 plant and fruit traits on over 3000 genotypes that were represented in either the current breeding population or as progenitors in the pedigree of these individuals. The resulting linear combination (i.e., sum of economic weight × breeding value for all 12 traits) estimated the overall economic worth of each genotype as H, the aggregate economic genotype. H values were validated by comparisons among commercial cultivars and were also compared with the estimated gross margins. When the H value of ‘Festival’ was set as zero, the H values of genotypes in the pedigree ranged from –0.36 to +0.28. H was highly correlated (R2 = 0.77) with the year of selection (1945–98). The gross margins were highly linearly related (R2 > 0.98) to H values when the genotype was planted on less than 50% of available area, but the relationship was non-linear [quadratic with a maximum (R2 > 0.96)] when the planted area exceeded 50%. Additionally, with H values above zero, the variation in gross margin increased with increasing H values as the percentage of area planted to a genotype increased. High correlations among some traits allowed the omission of any one of three of the 12 traits with little or no effect on ranking (Spearman’s rank correlation 0.98 or greater). Thus, these traits may be dropped from the aggregate economic genotype, leading to either cost reductions in the breeding program or increased selection intensities for the same resources. H was efficient in identifying economically superior genotypes for breeding and deployment, but because of the non-linear relationship with gross margin, calculation of a gross margin for genotypes with high H is also necessary when cultivars are deployed across more than 50% of the available area.