Genomic Reconstruction of the Domestication History of Sinningia speciosa (Lodd.) Hiern, and the Development of a Novel Genotyping Approach

• Main Author: Hasing Rodriguez, Tomas Nestor
• Other Authors: Horticulture
• Format: Others
• Published: Virginia Tech 2019
• Subjects: Phenotypic diversity; population structure; genetic bottleneck; ornamental crop; plant domestication; reduced representation
• Online Access: http://hdl.handle.net/10919/95510

Most staple food crops were domesticated thousands of years ago through independent processes across different regions of the world. Studies of the history of such crops have been essential to our understanding of plant domestication as a process that started with the collection of wild material and continued with subsequent propagation, cultivation, and selection under human care. Domestication often involves a complex genetic structure with contributions from multiple founder populations, interspecific hybridization, chromosomal introgressions, and polyploidization events that occurred hundreds to thousands of years earlier. Such intricate origins complicate the systematic study of the sources of phenotypic variation. The analysis of recently domesticated, non-traditional, non-model species, such as Sinningia speciosa (Gesneriaceae), can expand the knowledge that we have on phenotypic variation under domestication, and help us to comprehend modern patterns of plant domestication and to broaden our understanding of the general trends. S. speciosa is commonly known as the 'florist's gloxinia', and it has been cultivated for 200 years as an ornamental houseplant. In our genomic study of S. speciosa, we examined an extensive diversity panel consisting of 115 individuals that included different species in the genus, wild representatives, and cultivated accessions, as well as 150 individuals from an F2 segregating population. Our analyses revealed that all of the domesticated varieties are derived from a single founder population that originated in or near the city of Rio de Janeiro in Brazil. We identified two loci associated with domesticated traits (flower symmetry and color) and did not detect any major hybridization or polyploidization events that could have contributed to the rapid increase in phenotypic diversity. Our findings, in conjunction with other features such as a small, low-complexity genome, ease of cultivation, and rapid generation time, makes this species an attractive model for the study of genomic variation under domestication. Basic research on non-model organisms with low economic importance is uncommon but necessary to understand the world from a broader perspective. In such cases, reduced representation approaches like Genotyping-by-Sequencing (GBS) are efficient low-cost alternatives to whole genome resequencing. However, most of these technologies are subject to patent protection, licensing processes, and fees that constrain genomic research for small non-profit research organizations. We have designed a protocol to construct reduced representation libraries from genomic DNA. Our approach, called Targeted Amplification of Scattered Sites (TASS), deviates from the traditional digestion-ligation-amplification process that is the subject of intellectual property that protects most current methods. Instead, TASS relies on 1) targeting and duplicating scattered regions in the genome by annealing and expanding long tail primers with short annealing sites, and 2) amplifying these regions using primers that are complementary to the added overhangs. At the moment GBS is more consistent and delivers more variants than TASS. However, we have established a foundation on which further optimization can produce an accessible, easy to implement, high-throughput genotyping approach. === Doctor of Philosophy === Most staple food crops were domesticated thousands of years ago through unrelated processes that were initiated across different regions of the world. Studies of the history of such crops have been essential to our understanding of plant domestication, a process that started with the collection of wild material and continued with subsequent propagation and cultivation under human care. Plant domestication has often involved a complex combination of ancestral lineages that encompass multiple populations, crosses with other species, and large DNA reorganizations that occurred hundreds to thousands of years earlier. Such intricate origins make the systematic study of plant domestication very challenging. The analysis of recently domesticated plants such as the 'florist's gloxinia' (Sinningia speciosa), can help us to better understand some of the changes that have occurred during domestication, as well as to comprehend modern patterns of plant domestication and to broaden our understanding of general trends. Florist's gloxinias are ornamental plants that have been cultivated during the last 200 years. In this study we examined 115 specimens, including wild and cultivated types of florist's gloxinias, as well as closely related species in Sinningia. We also constructed and evaluated an artificial population of 150 individuals from the cross of a wild and a cultivated form. Our analyses revealed that all of the domesticated varieties are descendants from a single wild population that originated in or near the city of Rio de Janeiro in Brazil. We also identified two regions of DNA that are responsible for the changes in flower shape and color, and crosses with other species did not introduce such alterations. Our findings, in conjunction with other features such as its small nuclear DNA content, the ease of cultivation indoors, and a rapid generation time, makes the florists' gloxinia an attractive crop to the study the effects of plant domestication. Research on organisms with low economic importance is uncommon but necessary to understand the world from a broader perspective. In such cases, analyzing the entire genetic information that is stored as DNA may be cost-prohibitive. Instead, approaches that sample small portions of DNA from each individual can be utilized. Most of these technologies are currently patented and subject to licensing processes and fees that limit their implementation by small non-profit research organizations. In this study we designed a protocol to sample small portions of DNA, similarly to existing techniques. However, our approach, called Targeted Amplification of Scattered Sites (TASS), employs a sampling process that deviates from the traditional patented procedure that is used in most current methods. At present, TASS is not as consistent and delivers less information than traditional approaches. However, we have established a foundation on which further optimization can produce an accessible and easy to implement technique.