Complete chloroplast genomes of three copal trees (Bursera: Bullockia): comparative analysis and phylogenetic relationships.

BACKGROUND: Bursera trees are conspicuous elements of the tropical dry forests in the Neotropics that have significant cultural value due to their fragrant resins (incense), wood sources (handcrafts), and ecological benefits. Despite their relevance, genetic resources developed for the genus are scarce. METHODS AND RESULTS: We obtained the complete chloroplast (Cp) genome sequence, analyzed the genome structure, and performed functional annotation of three Bursera species of the Bullockia section: Bursera cuneata, B. palmeri, and B. bipinnata. The Cp genome sizes ranged from 159,824 to 159,872 bp in length, including a large single-copy (LSC) region from 87,668 to 87,656 bp, a small single-copy (SSC) from 18,581 to 18,571 bp, and two inverted repeats regions (IRa and IRb) of 26,814 bp each. The three Cp genomes consisted of 135 genes, of which 90 were protein-coding, 37 tRNAs, and 8 rRNAs. The Cp genomes were relatively conserved, with the LSC region exhibiting the greatest nucleotide divergence (psbJ, trnQ-UCC, trnG-UCC, and petL genes), whereas few changes were observed in the IR border regions. Between 589 and 591 simple sequence repeats were identified. Analysis of phylogenetic relationships using our data for each Cp region (LSC, SSC, IRa, and IRb) and of seven species within Burseraceae confirmed that Commiphora is the sister genus of Bursera. Only the phylogenetic trees based on the SSC and LSC regions resolved the close relationship between B. bipinnata and B. palmeri. CONCLUSION: Our work contributes to the development of Bursera's genomic resources for taxonomic, evolutionary, and ecological-genetic studies.

Development of Genomic Resources in Mexican Bursera (Section: Bullockia: Burseraceae): Genome Assembly, Annotation, and Marker Discovery for Three Copal Species.

Bursera comprises ~100 tropical shrub and tree species, with the center of the species diversification in Mexico. The genomic resources developed for the genus are scarce, and this has limited the study of the gene flow, local adaptation, and hybridization dynamics. In this study, based on ~155 million Illumina paired-end reads per species, we performed a de novo genome assembly and annotation of three Bursera species of the Bullockia section: Bursera bipinnata, Bursera cuneata, and Bursera palmeri. The total lengths of the genome assemblies were 253, 237, and 229 Mb for B. cuneata, B. palmeri, and B. bipinnata, respectively. The assembly of B. palmeri retrieved the most complete and single-copy BUSCOs (87.3%) relative to B. cuneata (86.5%) and B. bipinnata (76.6%). The ab initio gene prediction recognized between 21,000 and 32,000 protein-coding genes. Other genomic features, such as simple sequence repeats (SSRs), were also detected. Using the de novo genome assemblies as a reference, we identified single-nucleotide polymorphisms (SNPs) for a set of 43 Bursera individuals. Moreover, we mapped the filtered reads of each Bursera species against the chloroplast genomes of five Burseraceae species, obtaining consensus sequences ranging from 156 to 160 kb in length. Our work contributes to the generation of genomic resources for an important but understudied genus of tropical-dry-forest species.

Molecular evidence and ecological niche modeling reveal an extensive hybrid zone among three Bursera species (section Bullockia).

The genus Bursera, includes ~100 shrub and trees species in tropical dry forests with its center of diversification and endemism in Mexico. Morphologically intermediate individuals have commonly been observed in Mexican Bursera in areas where closely related species coexist. These individuals are assumed to result from interspecific hybridization, but no molecular evidence has supported their hybrid origins. This study aimed to investigate the existence of interspecific hybridization among three Mexican Bursera species (Bullockia section: B. cuneata, B. palmeri and B. bipinnata) from nine populations based on DNA sequences (three nuclear and four chloroplast regions) and ecological niche modeling for three past and two future scenario projections. Results from the only two polymorphic nuclear regions (PEPC, ETS) supported the hybrid origin of morphologically intermediate individuals and revealed that B. cuneata and B. bipinnata are the parental species that are genetically closer to the putative hybrids. Ecological niche modeling accurately predicted the occurrence of putative hybrid populations and showed a potential hybrid zone extending in a larger area (74,000 km2) than previously thought. Paleo-reconstructions showed a potential hybrid zone existing from the Last Glacial Maximum (~ 21 kya) that has increased since the late Holocene to the present. Future ecological niche projections show an increment of suitability of the potential hybrid zone for 2050 and 2070 relative to the present. Hybrid zone changes responded mostly to an increase in elevational ranges. Our study provides the first insight of an extensive hybrid zone among three Mexican Bursera species based on molecular data and ecological niche modeling.

Testing the hypothesis that biological modularity is shaped by adaptation: Xylem in the Bursera simaruba clade of tropical trees.

The study of modularity allows recognition of suites of character covariation that potentially diagnose units of evolutionary change. One prominent perspective predicts that natural selection should forge developmental units that maximize mutual functional independence. We examined the module-function relation using secondary xylem (wood) in a clade of tropical trees as a study system. Traditionally, the three main cell types in wood (vessels, fibers, and parenchyma) have respectively been associated with three functions (conduction, mechanical support, and storage). We collected samples from nine species of the simaruba clade of Bursera at fifteen sites and measured thirteen anatomical variables that have traditionally been regarded as reflecting the distinct functions of these cell types. If there are indeed (semi) independently evolving modules associated with functions, and cell types really are associated with these functions, then we should observe greater association between traits within cell types than between traits from different cell types. To map these associations, we calculated correlation coefficients among anatomical variables and identified modules using cluster and factor analysis. Our results were only partially congruent with expectations, with associations between characters of different cell types common. These results suggest causes of covariation, some involving selected function as predicted, but also highlighting the tradeoffs and shared developmental pathways limiting the evolutionary independence of some cell types in the secondary xylem. The evolution of diversity across the simaruba clade appears to have required only limited independence between parts.

Ontogenetic modulation of branch size, shape, and biomechanics produces diversity across habitats in the Bursera simaruba clade of tropical trees.

Organismal size and shape inseparably interact with tissue biomechanical properties. It is therefore essential to understand how size, shape, and biomechanics interact in ontogeny to produce morphological diversity. We estimated within species branch length-diameter allometries and reconstructed the rates of ontogenetic change along the stem in mechanical properties across the simaruba clade in the tropical tree genus Bursera, measuring 376 segments from 97 branches in nine species in neotropical dry to rain forest. In general, species with stiffer materials had longer, thinner branches, which became stiffer more quickly in ontogeny than their counterparts with more flexible materials. We found a trend from short stature and flexible tissues to tall statures and stiff tissues across an environmental gradient of increasing water availability, likely reflecting a water storage-mechanical support tradeoff. Ontogenetic variation in size, shape, and mechanics results in diversity of habits, for example, rapid length extension, sluggish diameter expansion, and flexible tissues results in a liana, as in Bursera instabilis. Even species of similar habit exhibited notable changes in tissue mechanical properties with increasing size, illustrating the inseparable relationship between organismal proportions and their tissue mechanics in the ontogeny and evolution of morphological diversity.

Differential effects of landscape-level environmental features on genetic structure in three codistributed tree species in Central America.

Landscape genetic studies use spatially explicit population genetic information to determine the physical and environmental causes of population genetic structure on regional scales. Comparative studies that identify common barriers to gene flow across multiple species within a community are important to both understand the evolutionary trajectories of populations and prioritize habitat conservation. Here, we use a comparative landscape genetic approach to ask whether gradients in temperature or precipitation seasonality structure genetic variation across three codistributed tree species in Central America, or whether a simpler (geographic distance) or more complex, species-specific environmental niche model is necessary to individually explain population genetic structure. Using descriptive statistics and causal modelling, we find that different factors best explain genetic distance in each of the three species: environmental niche distance in Bursera simaruba, geographic distance in Ficus insipida and historical barriers to gene flow or cryptic reproductive barriers for Brosimum alicastrum. This study confirms suggestions from previous studies of Central American tree species that imply that population genetic structure of trees in this region is determined by complex interactions of both historical and current barriers to gene flow.

Insights into the historical construction of species-rich Mesoamerican seasonally dry tropical forests: the diversification of Bursera (Burseraceae, Sapindales).

• Mesoamerican arid biomes epitomize neotropical rich and complex biodiversity. To document some of the macroevolutionary processes underlying the vast species richness of Mesoamerican seasonally dry tropical forests (SDTFs), and to evaluate specific predictions about the age, geographical structure and niche conservatism of SDTF-centered woody plant lineages, the diversification of Bursera is reconstructed. • Using a nearly complete Bursera species-level phylogeny from nuclear and plastid genomic markers, we estimate divergence times, test for phylogenetic and temporal diversification heterogeneity, test for geographical structure, and reconstruct habitat shifts. • Bursera became differentiated in the earliest Eocene, but diversified during independent early Miocene consecutive radiations that took place in SDTFs. The late Miocene average age of Bursera species, the presence of phylogenetic geographical structure, and its strong conservatism to SDTFs conform to expectations derived from South American SDTF-centered lineages. • The diversification of Bursera suggests that Mesoamerican SDTF richness derives from high speciation from the Miocene onwards uncoupled from habitat shifts, during a period of enhanced aridity resulting mainly from global cooling and regional rain shadows.

Diversification in species complexes: tests of species origin and delimitation in the Bursera simaruba clade of tropical trees (Burseraceae).

Molecular phylogenies are invaluable for testing morphology-based species delimitation in species complexes, as well as for examining hypotheses regarding the origination of species in these groups. Using five nucleotide markers, we reconstructed the phylogeny of the Bursera simaruba species complex of neotropical trees to test the notion that four "satellite" species originated from populations of the most widely distributed member of the genus, B. simaruba, which the satellites strongly resemble. In addition to molecular phylogenetic reconstruction, we tested species delimitation of B. simaruba and the satellites using multivariate analyses of morphological and ecological characters. The analyses evaluated the taxonomic value of these traditional characters and pinpointed those in need of further study, such as the expression of pubescence. Phylogenetic data rejected the origin of three satellite species from their purported ancestor, B. simaruba, and we ascribe their morphological similarity to convergence or parallelism. The fourth satellite species likely represents one end of a spectrum of inflorescence length variation within B. simaruba and is conspecific. Despite its marked morphological variability, we recovered B. simaruba as a single valid species, which implies that it maintains genetic cohesion among distant populations throughout its vast range.

Macroevolutionary chemical escalation in an ancient plant-herbivore arms race.

A central paradigm in the field of plant-herbivore interactions is that the diversity and complexity of secondary compounds in plants have intensified over evolutionary time, resulting in the great variety of secondary products that currently exists. Unfortunately, testing of this proposal has been very limited. We analyzed the volatile chemistry of 70 species of the tropical plant genus Bursera and used a molecular phylogeny to test whether the species' chemical diversity or complexity have escalated. The results confirm that as new species diverged over time they tended to be armed not only with more compounds/species, but also with compounds that could potentially be more difficult for herbivores to adapt to because they belong to an increasing variety of chemical pathways. Overall chemical diversity in the genus also increased, but not as fast as species diversity, possibly because of allopatric species gaining improved defense with compounds that are new locally, but already in existence elsewhere.

Sources and sinks of diversification and conservation priorities for the Mexican tropical dry forest.

Elucidating the geographical history of diversification is critical for inferring where future diversification may occur and thus could be a valuable aid in determining conservation priorities. However, it has been difficult to recognize areas with a higher likelihood of promoting diversification. We reconstructed centres of origin of lineages and identified areas in the Mexican tropical dry forest that have been important centres of diversification (sources) and areas where species are maintained but where diversification is less likely to occur (diversity sinks). We used a molecular phylogeny of the genus Bursera, a dominant member of the forest, along with information on current species distributions. Results indicate that vast areas of the forest have historically functioned as diversity sinks, generating few or no extant Bursera lineages. Only a few areas have functioned as major engines of diversification. Long-term preservation of biodiversity may be promoted by incorporation of such knowledge in decision-making.

The impact of herbivore-plant coevolution on plant community structure.

Coevolutionary theory proposes that the diversity of chemical structures found in plants is, in large part, the result of selection by herbivores. Because herbivores often feed on chemically similar plants, they should impose selective pressures on plants to diverge chemically or bias community assembly toward chemical divergence. Using a coevolved interaction between a group of chrysomelid beetles and their host plants, I tested whether coexisting plants of the Mexican tropical dry forest tend to be chemically more dissimilar than random. Results show that some of the communities are chemically overdispersed and that overdispersion is related to the tightness of the interaction between plants and herbivores and the spatial scale at which communities are measured. As coevolutionary specialization increases and spatial scale decreases, communities tend to be more chemically dissimilar. At fairly local scales and where herbivores have tight, one-to-one interactions with plants, communities have a strong pattern of chemical disparity.

Timing the origin and expansion of the Mexican tropical dry forest.

Macroevolution examines the temporal patterns of biological diversity in deep time. When combined with biogeography, it can provide unique information about the historical changes in the distribution of communities and biomes. Here I document temporal and spatial changes of diversity in the genus Bursera and relate them to the origin and expansion of the tropical dry forests of Mexico. Bursera is very old, highly adapted to warm dry conditions, and a dominant member of the Mexican tropical dry forest. These characteristics make it a useful indicator of the history of this vegetation. I used a time-calibrated phylogeny to estimate Bursera's diversification rate at different times over the last 60 million years. I also reconstructed the geographic center and time of origin of all species and nodes from information on current distributions. Results show that between 30 and 20 million years ago, Bursera began a relatively rapid diversification. This suggests that conditions were favorable for its radiation and thus, very probably for the establishment of the dry forest as well. The oldest lineages diverged mostly in Western Mexico, whereas the more recent lineages diverged in the south-central part of the country. This suggests that the tropical dry forest probably first established in the west and then expanded south and east. The timing of the radiations in these areas corresponds to that suggested for formations of the mountainous systems in Western and Central Mexico, which have been previously recognized as critical for the persistence of the Mexican dry forest.

Evolution of Mexican Bursera (Burseraceae) inferred from ITS, ETS, and 5S nuclear ribosomal DNA sequences.

I reconstructed a phylogeny of 66 species and varieties of Bursera and 9 outgroup species using sequences of the internal transcribed spacer region (ITS), the 5S non-transcribed region (5S-NTS), and the external transcribed region (ETS) of nuclear ribosomal DNA. This study extends a previously proposed parsimony-based phylogenetic study that used the ITS sequences of 57 Bursera species and five outgroups. Parsimony and maximum likelihood methods were used to infer the phylogeny in this new study. Analyses of the combined data sets largely confirmed the phylogenetic relationships proposed by the previous molecular study but generated a considerably more robust topology. The new phylogenies corroborate the monophyly of the genus, and its division into the two monophyletic subgenera or sections, Bursera and Bullockia. The current analyses also identify four main groups of species in section Bursera, and two in section Bullockia, confirming some of the previously proposed groups based on fruit, flower, and leaf morphology. One previously problematic species B. sarcopoda, which has sometimes been placed in Commiphora, is shown to belong in Bursera. Another controversial species, Commiphora leptophloeos, which was thought to belong to Bursera, falls within Commiphora.