Juan Carlos Motamayor

This study sought to quantify and characterize diverse rootstock scion interactions in cacao around graft compatibility, disease resistance, nutrient use efficiency, vigor traits, and translocation of nonstructural carbohydrates. In total, 106 grafts were performed with three scion cultivars (Matina 1/6, Criollo 22, Pound 7) and nine diverse open-pollinated seedling populations (BYNC, EQX 3348, GNV 360, IMC 14, PA 107, SCA 6, T 294, T 384, T 484). We found evidence for both local and translocated graft incompatibility. Cross sections and Micro-XCT imaging revealed anatomical anomalies, including necrosis and cavitation at the junction and accumulation of starch in the rootstock directly below the graft junction. Scion genetics were a significant factor in explaining differences in graft take, and graft take varied from 47% (Criollo 22) to 72% (Pound 7). Rootstock and scion identity both accounted for differences in survival over the course of the 30-month greenhouse study, with a low of 28.5% survival of Criollo 22 scions and a high of 72% for Pound 7 scions. Survival by rootstocks varied from 14.3% on GNV 360 to 100% survival on T 294 rootstock. A positive correlation of 0.34 (p = 0.098) was found between the graft success of different rootstock–scion combinations and their kinship coefficient, suggesting that relatedness of stock and scion could be a driver of incompatibility. Significant rootstock–scion effects were also observed for nutrient use efficiency, plant vigor, and resistance to Phytophthora palmivora. These findings, while preliminary in nature, highlight the potential of rootstock breeding to improve plant nutrition, resilience, and disease resistance in cacao.

Predicting the potential fate of a species in the face of climate change requires knowing the distribution of molecular adaptations across the geographic range of the species. In this work, we analyzed 79 genomes of Theobroma cacao, an Amazonian tree known for the fruit from which chocolate is produced, to evaluate how local and regional molecular signatures of adaptation are distributed across the natural range of the species. We implemented novel techniques that incorporate summary statistics from multiple selection scans to infer selective sweeps. The majority of the molecular adaptations in the genome are not shared among populations. We show that ~71.5% of genes under selection also show significant associations with changes in environmental variables. Our results support the interpretation that these genes contribute to local adaptation of the populations in response to abiotic factors. We also found strong patterns of molecular adaptation in a diverse array of disease resistance genes (6.5% of selective sweeps), suggesting that differential adaptation to pathogens also contributes significantly to local adaptations. Our results are consistent with the interpretation that local selective pressures are more important than regional selective pressures in explaining adaptation across the range of a species.

Domestication has had a strong impact on the development of modern societies. We sequenced 200 genomes of the chocolate plant Theobroma cacao L. to show for the first time to our knowledge that a single population, the Criollo population, underwent strong domestication ~3600 years ago (95% CI: 2481–13,806 years ago). We also show that during the process of domestication, there was strong selection for genes involved in the metabolism of the colored protectants anthocyanins and the stimulant theobromine, as well as disease resistance genes. Our analyses show that domesticated populations of T. cacao (Criollo) maintain a higher proportion of high-frequency deleterious mutations. We also show for the first time the negative consequences of the increased accumulation of deleterious mutations during domestication on the fitness of individuals (significant reduction in kilograms of beans per hectare per year as Criollo ancestry increases, as estimated from a GLM, P = 0.000425).

Corrigendum: A larger chocolate chip-development of a 15K theobroma cacao L. SNP array to create high-density linkage maps.Name of all authors as they appear in the published original article (If correcting author names please put the correct spelling. If adding authors, please include additional authors.)Donald Livingstone III1†, Conrad Stack1†, Guiliana M. Mustiga1, Dayana C. Rodezno1, Carmen Suarez2,3, Freddy Amores2,4, Frank A. Feltus5, Keithanne Mockaitis6, Omar E. Cornejo7 and Juan C. Motamayor1*Affiliations of all authors as they appear in the published original version of the article 1Mars, Inc., Miami, FL, United States 2Estación Experimental Tropical Pichilingue, National Institute of Agricultural Research, Quevedo, Ecuador 3Agricultural Faculty, Technical University of Quevedo, Quevedo, Ecuador 4Universidad Técnica Estatal Quevedo (UTEQ), Quevedo, Ecuador 5Department of Genetics and Biochemistry, Clemson University, Clemson, SC, United States 6Department of Biology and Pervasive Technology Institute, Indiana University Bloomington, Bloomington, IN, United States 7School of Biological Sciences, Washington State University, Pullman, WA, United States* Correspondence: email of the corresponding author from the original version of the articleKeywords: cacao, SNP, genotyping, QTLCorrigendum on: add full citation of the original version of the article. Livingstone, D., Stack, C., Mustiga, G. M., Rodezno, D. C., Suarez, C., Amores, F., et al. (2017). A larger chocolate chip-development of a 15K theobroma cacao L. SNP array to create high-density linkage maps. Front. Plant Sci. 8. doi:10.3389/fpls.2017.02008.Error in Figure/TableIn the original article, there was a mistake in Supplemental Table 2 as published online. Supplemental Table 2 is a duplication of Supplemental Table 3. The corrected Supplemental Table 2 appears below and is attached as separate excel file for convenience. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way.

Theobroma cacao L. is a tropical tree originating in the Amazon, where it grows naturally in the shade of tropical rainforests. Cacao sub-products, such as butter and powder, are produced as principal components of chocolate and contain important nutritional compounds such as polyphenols and flavonoids. However, bean production is decreasing because plantations are antiquated and unproductive. Cacao propagation has been traditionally performed through classical propagation methods, such as grafting or rooted cuttings, but those methods are not sufficient to obtain large quantities of planting material with the desired genetic quality and optimal plant health. In the search for solutions to this problem, somatic embryogenesis (SE) is a vegetative method used for cacao propagation that has the potential to be explored. SE is a type of clonal propagation by which totipotent cells in the somatic tissue can develop into embryos and subsequently convert into plants. This method offers significant technological advantages because it is possible to obtain a large quantity of disease-free planting material with good agronomic characteristics and genetic stability. In T. cacao, tow techniques of in vitro micropropagation have been reported as direct and indirect SE. Indirect SE requires the additional step of cell dedifferentiation, unlike direct SE, which does not require this step. Here, we report a protocol using direct and indirect SE techniques using two types of culture methodologies—solid and liquid culture media.

A comprehensive understanding of the genetic basis of target traits in any crop is critical to design breeding strategies for the development and release of new improved varieties. In this study, 34 cacao families were evaluated for vigor and yield related traits over the course of 6 years in Costa Rica. Linear mixed models provided the variance components for the partitioning of additive and non-additive effects. Heritabilities of yield over time ranged from 0.085 to 0.576, from 0.127 to 0.399 for vigor, and 0.141 to 0.146 for disease resistance traits. Significant (p < 0.001) general combining abilities were observed for ICS-43 and LcTeen-37 with negative effect on average yield (−0.674, −0.690), respectively. Specific combining abilities for yield had significant (p < 0.001) positive effect from the cross GU-154-L x UF-273 Type 2 (0.703) and strong negative interaction between ICS-43 and LF-1 (−0.975). A weighted index was used to select the top performers while providing the corresponding genetic gains. At an 1% selection intensity, yield component gains ranged from 17.8 to 331.9%. Agronomic traits such as branch angle, trunk diameter and jorquette height had lower genetic gains and lower heritabilities. In addition, the parents in this study were genotyped with a 96-SNP marker off-typing set and a significant positive correlation of 0.39 (p = 0.019) was found between genetic distance and specific combining abilities for yield. Preliminary comparison of clonal parents vs. seedlings yield in the family with the highest SCA suggest for the first time presence of heterobeltiosis in cacao.

Cacao (Theobroma cacao) is a globally important crop, and its yield is severely restricted by disease. Two of the most damaging diseases, witches’ broom disease (WBD) and frosty pod rot disease (FPRD), are caused by a pair of related fungi: Moniliophthora perniciosa and Moniliophthora roreri, respectively. Resistant cultivars are the most effective long-term strategy to address Moniliophthora diseases, but efficiently generating resistant and productive new cultivars will require robust methods for screening germplasm before field testing. Marker-assisted selection (MAS) and genomic selection (GS) provide two potential avenues for predicting the performance of new genotypes, potentially increasing the selection gain per unit time. To test the effectiveness of these two approaches, we performed a genome-wide association study (GWAS) and GS on three related populations of cacao in Ecuador genotyped with a 15K single nucleotide polymorphism (SNP) microarray for three measures of WBD infection (vegetative broom, cushion broom, and chirimoya pod), one of FPRD (monilia pod) and two productivity traits (total fresh weight of pods and % healthy pods produced). GWAS yielded several SNPs associated with disease resistance in each population, but none were significantly correlated with the same trait in other populations. Genomic selection, using one population as a training set to estimate the phenotypes of the remaining two (composed of different families), varied among traits, from a mean prediction accuracy of 0.46 (vegetative broom) to 0.15 (monilia pod), and varied between training populations. Simulations demonstrated that selecting seedlings using GWAS markers alone generates no improvement over selecting at random, but that GS improves the selection process significantly. Our results suggest that the GWAS markers discovered here are not sufficiently predictive across diverse germplasm to be useful for MAS, but that using all markers in a GS framework holds substantial promise in accelerating disease-resistance in cacao.

Cacao is an important crop, its beans are key raw materials for the chocolate and cosmetic industries. Ceratocystis wilt of cacao (CWC) caused by Ceratocystis cacaofunesta is a lethal disease for the crop. Therefore, the selection of resistant cacao varieties is one of the viable ways to minimize losses in cacao production. In this paper, we described the identification of a major QTL associated with CWC in an F1 mapping population from a cross between a resistant, “TSH 1188,” and a susceptible genotype, “CCN 51.” A set of 266 trees were genotyped using 3,526 single nucleotide polymorphic markers and then multiple QTL mapping analyses were performed. Two QTLs were identified on chromosomes IV and VI. The major QTL was located at 20 cM from the top position of chromosome VI, accounting for more than 60% of the phenotypic variation. The favorable allele T1, with haplotype GTT, came from the “TSH 1188” parent. It was evident that the haplotype combination T1C2 on chromosome VI was the most significant for resistance, since 93% of resistant trees had this haplotype. The major QTL converged to a genomic region of 739.4 kb that harbored nine candidate genes, including two major classes of resistance genes, which would make them the primary candidates involved in the resistance to CWC. The haplotypes detected are now used to improve the efficiency and precision of the selection of resistant trees in cacao breeding.

Cacao (Theobroma cacao L.) is an important cash crop in tropical regions around the world and has a rich agronomic history in South America. As a key component in the cosmetic and confectionary industries, millions of people worldwide use products made from cacao, ranging from shampoo to chocolate. An Illumina Infinity II array was created using 13,530 SNPs identified within a small diversity panel of cacao. Of these SNPs, 12,643 derive from variation within annotated cacao genes. The genotypes of 3,072 trees were obtained, including two mapping populations from Ecuador. High-density linkage maps for these two populations were generated and compared to the cacao genome assembly. Phenotypic data from these populations were combined with the linkage maps to identify the QTLs for yield and disease resistance.