Numerous collecting expeditions of Theobroma cacao L. germplasm have been undertaken in Latin-America. However, most of this germplasm has not contributed to cacao improvement because its relationship to cultivated selections was poorly understood. Germplasm labeling errors have impeded breeding and confounded the interpretation of diversity analyses. To improve the understanding of the origin, classification, and population differentiation within the species, 1241 accessions covering a large geographic sampling were genotyped with 106 microsatellite markers. After discarding mislabeled samples, 10 genetic clusters, as opposed to the two genetic groups traditionally recognized within T. cacao, were found by applying Bayesian statistics. This leads us to propose a new classification of the cacao germplasm that will enhance its management. The results also provide new insights into the diversification of Amazon species in general, with the pattern of differentiation of the populations studied supporting the palaeoarches hypothesis of species diversification. The origin of the traditional cacao cultivars is also enlightened in this study.

Hydrofluorocarbons, currently used as refrigerants in air-conditioning systems, are potent greenhouse gases, and their contribution to climate change is projected to increase. Future use of the hydrofluorocarbons will be phased down and, thus replacement fluids must be found. Here we show that only a few pure fluids possess the combination of chemical, environmental, thermodynamic, and safety properties necessary for a refrigerant and that these fluids are at least slightly flammable. We search for replacements by applying screening criteria to a comprehensive chemical database. For the fluids passing the thermodynamic and environmental screens (critical temperature and global warming potential), we simulate performance in small air-conditioning systems, including optimization of the heat exchangers. We show that the efficiency-versus-capacity trade-off that exists in an ideal analysis disappears when a more realistic system is considered. The maximum efficiency occurs at a relatively high volumetric refrigeration capacity, but there are few fluids in this range.

A novel liquid chromatographic system which enables high temperature ultrafast liquid chromatography (HTU-FLC) has been designed through the careful consideration of heat transfer, band broadening, and pressure drop. Studies of the effect of linear velocity on the HETP show that column efficiency at high velocity, especially of well-retained solutes, dramatically improves at higher temperatures. At 150 degrees C, at a flow rate of 15 mL/min with a 5 cm by 4.6 mm (i.d.) column packed with 3 microns polystyrene-coated zirconia porous particles, long chain alkylphenones were completely resolved, and the analysis time could be decreased by a factor of 50 compared to that at room temperature (25 degrees C) at a conventional flow rate (4 mL/min). In addition, using pure water as the mobile phase, five phenols were separated in less than 30 s.

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There are now available a variety of tunable solvents; these have been used extensively for extractions and in a variety of materials applications. Our focus has been to apply these techniques to chemical reactions to take advantage of the special properties available, primarily for sustainable technology, to create processes that are potentially more benign and more advantageous. We report here our work in using supercritical fluids, near-critical fluids, and gas-expanded liquids to couple organic reactions with separations. In this paper, we review applications involving improved transport, catalyst recycling, and product separation as well as the in situ generation of catalysts. Although such tunable solvents are in no way a panacea, they do offer the chemical community alternatives that can often be applied creatively to many reaction opportunities.

We monitored PCR in volumes of the order of 10 nl in glass microcapillaries using a fluorescence energy transfer assay in which fluorescence increases if product is made due to template-dependent nucleolytic degradation of an internally quenched probe (TaqMan assay). This assay detected single starting template molecules in dilutions of genomic DNA. The results suggest that it may be feasible to determine the number of template molecules in a sample by counting the number of positive PCRs in a set of replicate reactions using terminally diluted sample. Since the assay system is closed and potentially automatable, it has promise for clinical applications.

Three horticultural races of avocado (Persea americana Mill.) are known: Guatemalan, Mexican, and West Indian. Each race has unique characteristics and current commercial varieties have been selected from within the races or from interracial hybrids. Using 14 microsatellite loci we investigated the genetic variation among 224 accessions (394 plants) maintained at the National Germplasm Repository (NGR) in Miami, Fla., and a set of 34 clones from the University of California South Coast Field Station (SCFS) located in Irvine, Calif. The 14 microsatellite loci had an average of 18.8 alleles per locus and average unbiased genetic diversity was 0.83. The total propagation error in the collection, i.e., plants that had been incorrectly labeled or grafted, was estimated to be 7.0%. Although many unique alleles did exist, no useful race-specific markers were found. A general concordance between the horticultural race and the clusters obtained from molecular data was observed. Principal Coordinate Analysis (PCA) grouped the Guatemalan and Mexican races into two distinct clusters. The West Indian also grouped into a unique major cluster but with an outlying group. Using the PCA a change in the racial designation or interracial hybrid status for 50 accessions (19.7%) is proposed. The unbiased gene diversity estimate was highest in the Mexican and Guatemalan races and lower in the West Indian group. This demonstrates the need to collect more of the West Indian germplasm to broaden the genetic diversity and to emphasize the identification of individuals conferring resistance to Phytophthora Root Rot (PRR).

A genetic linkage map was created from 146 cacao trees (Theobroma cacao), using an F2 population produced by selfing an F1 progeny of the cross Sca6 and ICS1. Simple sequence repeat (SSR) markers (170) were used principally for this map, with 12 candidate genes [eight resistance gene homologues (RGH) and four stress related WRKY genes], for a total of 182 markers. Joinmap software was used to create the map, and 10 linkage groups were clearly obtained, corresponding to the 10 known chromosomes of cacao. Our map encompassed 671.9 cM, approximately 100 cM less than most previously reported cacao maps, and 213.5 cM less than the one reported high-density map. Approximately 27% of the markers showed significant segregation distortion, mapping together in six genomic areas, four of which also showed distortion in other cacao maps. Two quantitative trait loci (QTL) for resistance to witches' broom disease were found, one producing a major effect and one a minor effect, both showing important dominance effects. One QTL for trunk diameter was found at a point 10.2 cM away from the stronger resistance gene. One RGH flanked the minor QTL for witches' broom resistance, implying possible association. QTLs mapped in F2 populations produce estimates of additive and dominance effects, not obtainable in F1 crosses. As dominance was clearly shown in the QTL found in this study, this population merits further study for evaluation of dominance effects for other traits. This F2 cacao population constitutes a useful link for genomic studies between cacao and cotton, its only widely grown agronomic relative.