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R.J. Schnell, C.T. Olano, J.S. Brown, A.W. Meerow, C. Cervantes-Martinez, C. Nagai and J.C. Motamayor

Commercial production of cacao in Hawaii is increasing, and this trend is expected to continue over the next several years. The increased acreages are being planted with seedlings from introduced and uncharacterized cacao populations from at least three initial introductions of cacao into the islands. Productive seedlings have been selected from a planting at Waialua, Oahu. The parents of these selections were believed to be the population at the Hawaii Agriculture Research Center (HARC) at Kunia; however, potential parental populations also exist at Univ. of Hawaii research stations at Waimanalo and Malama Ki. Using microsatellite markers, we analyzed the potential parental populations to identify the parents and determine the genetic background for 99 productive and 50 unproductive seedlings from the Waialua site. Based on 19 polymorphic microsatellite loci the parental population was identified as trees from Waimanalo and not trees from Malama Ki or Kunia. The Kunia and Malama Ki populations were very similar with low allelic diversity (A = 1.92) and low unbiased gene diversity (Hnb) of 0.311 and 0.329, respectively, and were determined to be Trinitario in type. The Waimanalo, productive seedling, and unproductive seedling populations had much higher levels of genetic diversity with Hnb of 0.699, 0.686, and 0.686, respectively, and were determined to be upper Amazon Forastero hybridized with Trinitario in type. An additional 46 microsatellite markers were amplified and analyzed in the Waimanalo parents, productive, and unproductive seedlings for a total of 65 loci. Seventeen loci contained alleles that were significantly associated with productive seedlings as determined by Armitage's trend test. Of these, 13 loci (76.4%) co-located with previously reported quantitative trait loci for productivity traits. These markers may prove useful for marker assisted selection and demonstrate the potential of association genetic studies in perennial tree crops such as cacao.

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J. Steven Brown, R.J. Schnell, J.C. Motamayor, Uilson Lopes, David N. Kuhn and James W. Borrone

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.

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R.J. Schnell, J.S. Brown, C.T. Olano, E.J. Power, C.A. Krol, D.N. Kuhn and J.C. Motamayor

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).

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Cuauhtemoc Cervantes-Martinez, J. Steven Brown, Raymond J. Schnell, Wilbert Phillips-Mora, Jemmy F. Takrama and Juan C. Motamayor

Knowledge of genetic differences among commonly cultivated cacao clones, as well as the type of gene action involved for disease resistance, yield, quality, and horticultural traits, are essential for cacao breeders to select parental clones efficiently and effectively. This information is also critical for quantitative geneticists in designing and improving quantitative trait loci (QTL) localization strategies using breeding populations, whether they involve analysis of multiple populations crossed to one common parent or association genetic analysis. The objectives of this research were to 1) verify the genetic identity of parental cacao clones used to produce hybrids for field evaluation at the Centro Agrónomico Tropical de Investigación y Enzeñanza (CATIE), Turrialba, Costa Rica, using molecular marker analysis, and 2) estimate general and specific combining ability (GCA and SCA) of the parental clones for resistance to frosty pod (Moniliophthora roreri Cif. and Par.) and black pod [Phytophthora palmivora (Butl.) Butl.] diseases, total number of pods, vigor (as measured by trunk diameter), and measures of maturity (months to first flowering and pod production). Misidentification of cacao clones was found at three levels. Molecular marker analysis revealed that six parental clones differed in identity to supposedly identical accessions from other germplasm collections. Trees of the parental clone UF 273 consisted of two clearly different genotypes, resulting in two types of progeny, requiring separate designation for correct statistical analysis. Out-crossed progeny, presumably from foreign pollen, and selfed progeny were also found. Two of the traits measured, percent healthy pods and percent pods with frosty pod, showed predominantly additive gene action, while the traits total number of pods and trunk diameter, demonstrated regulation by both additive and nonadditive gene action. Number of months to first flowering and first fruit both showed evidence of predominant regulation by nonadditive gene effects. Crosses of two parental clones, UF 712 and UF 273 Type I, were identified as potential candidates for QTL analysis as breeding populations, given their favorable GCA estimates for frosty pod resistance and total pod production, respectively.