Rebecca C. Jones

The genome of Eucalyptus grandis

Eucalypts are the world’s most widely planted hardwood trees. Their outstanding diversity, adaptability and growth have made them a global renewable resource of fibre and energy. We sequenced and assembled >94% of the 640-megabase genome of Eucalyptus grandis. Of 36,376 predicted protein-coding genes, 34% occur in tandem duplications, the largest proportion thus far in plant genomes. Eucalyptus also shows the highest diversity of genes for specialized metabolites such as terpenes that act as chemical defence and provide unique pharmaceutical oils. Genome sequencing of the E. grandis sister species E. globulus and a set of inbred E. grandis tree genomes reveals dynamic genome evolution and hotspots of inbreeding depression. The E. grandis genome is the first reference for the eudicot order Myrtales and is placed here sister to the eurosids. This resource expands our understanding of the unique biology of large woody perennials and provides a powerful tool to accelerate comparative biology, breeding and biotechnology.

A comparative analysis of population structure of a forest tree, Eucalyptus globulus (Myrtaceae), using microsatellite markers and quantitative traits

Eucalyptus globulus (Myrtaceae) is a forest tree native to southeastern Australia, but is grown globally for pulpwood and timber. Eight microsatellite loci were used to determine the degree of selectively neutral differentiation between native populations of the geographic races of E. globulus that are used in a national breeding programme. Spatial differentiation was detected among 340 samples from across the species range (FST=0.09±0.02). Analysis of molecular variance showed that there was significant variation between the races, and an unweighted pair group method with arithmetic mean analysis of Nei’s genetic distance between races showed that geographically proximal races tended to be more closely related than geographically distant races. This contrasted markedly with analyses based on quantitative genetic data, where some races appeared to be highly divergent from their geographically closest neighbours. Comparison of racial differentiation based on quantitative (QST) and molecular (FST) data suggested that at least five of the quantitative traits used for defining races of E. globulus have been influenced by natural selection, resulting in cases of both phenotypic divergence of parapatric races and phenotypic convergence of allopatric races. We conclude that selectively neutral molecular markers are more useful than quantitative genetic data for identifying the evolutionary affinities and lineages within E. globulus. However, both sources of information should be used in defining evolutionarily important units for conservation. The population structure observed in E. globulus has important consequences for future association studies and may also affect breeding strategies if significant genome co-adaptation has occurred.

Dispersal limitations, rather than bottlenecks or habitat specificity, can restrict the distribution of rare and endemic rainforest trees

Despite their narrow distribution, Australian rainforests still contain considerable levels of diversity and include many ancient, but often rare, lineages. Very little is known about the general biology of rainforest species, yet their long-term management depends on a better understanding of the main factors leading to rarity. For instance, are they highly endemic taxa, at the early stages of expansion, nearing the end of a period of decline, or persisting at low numbers over the long term? In this study we combine molecular, environmental, and ecological data to identify the factors responsible for the narrow distribution of a paleoendemic rainforest tree: Elaeocarpus sedentarius (Elaeocarpaceae). Between-population and between-generation comparisons of genetic diversity across all known populations of E. sedentarius show evidence of mutation-drift equilibrium rather than evidence of a recent bottleneck. Similarly, floristic and environmental data negate the hypothesis of rarity as a consequence of highly specialized habitat requirements. Instead, genetic structure and the available ecological data support the hypothesis of dispersal limitation as the main cause of endemism and that the species may have attained genetic equilibrium without realizing its full niche potential. We suggest that these factors are likely to explain narrow endemism in a broader range of taxa.

Multiple evolutionary processes drive the patterns of genetic differentiation in a forest tree species complex

Forest trees frequently form species complexes, complicating taxonomic classification and gene pool management. This is certainly the case in Eucalyptus, and well exemplified by the Eucalyptus globulus complex. This ecologically and economically significant complex comprises four taxa (sspp. bicostata, globulus, maidenii, pseudoglobulus) that are geographically and morphologically distinct, but linked by extensive “intergrade” populations. To resolve their genetic affinities, nine microsatellites were used to genotype 1200 trees from throughout the natural range of the complex in Australia, representing 33 morphological core and intergrade populations. There was significant spatial genetic structure (FST = 0.10), but variation was continuous. High genetic diversity in southern ssp. maidenii indicates that this region is the center of origin. Genetic diversity decreases and population differentiation increases with distance from this area, suggesting that drift is a major evolutionary process. Many of the intergrade populations, along with other populations morphologically classified as ssp. pseudoglobulus or ssp. globulus, belong to a “cryptic genetic entity” that is genetically and geographically intermediate between core ssp. bicostata, ssp. maidenii, and ssp. globulus. Geography, rather than morphology, therefore, is the best predictor of overall genetic affinities within the complex and should be used to classify germplasm into management units for conservation and breeding purposes.

Genetic diversity and mating system of an endangered tree Eucalyptus morrisbyi

Eucalyptus morrisbyi is an endangered eucalypt, restricted to four populations on the island of Tasmania. The two main populations are separated by 20 km, occurring in the Risdon Hills and on Calverts Hill, and differ markedly in size and health. Although they are both in reserves, the small population at Risdon Hills has experienced a marked decline in the last two decades. The other two populations (Lumeah Point and Honeywood Drive) are very small and under threat because of urbanisation. They are close to the large Calverts Hill population and may be remnants of a once larger population in this area prior to clearing for agriculture in the 19th century. A hypervariable chloroplast marker and six nuclear microsatellites, used to quantify genetic diversity among and within populations, indicated marked genetic differences between the two main populations (Risdon and Calverts Hills), with virtually no sharing of chloroplast haplotypes and little sharing of microsatellite alleles among populations. Both of the main populations are clearly required to adequately conserve the genetic diversity in this species, whereas the Lumeah Point and Honeywood Drive populations are similar to the proximal Calverts Hill population. The two main populations showed equally high levels of genetic diversity (average HE = 0.69) in the adult trees, using microsatellites, and little difference in inbreeding levels despite the large difference in population size. Analyses of 366 offspring from 9-11 trees from each main population revealed high outcrossing rates, little biparental inbreeding and high genetic diversity (average HE = 0.65) in both seedling populations. This indicates that open-pollinated seed collections from these populations capture sufficient genetic diversity for ex situ conservation plantings. It is argued that the high genetic diversity maintained in the small Risdon Hills population is due to a combination of the longevity of its genotypes (possibly up to 1155-1523 years) through a well developed mechanism of vegetative regeneration from lignotubers, coupled with high outcrossing rates maintained by a strong self-incompatibility mechanism.

Genetic Control of Heterochrony in Eucalyptus globulus

A change in the timing or rate of developmental events throughout ontogeny is referred to as heterochrony, and it is a major evolutionary process in plants and animals. We investigated the genetic basis for natural variation in the timing of vegetative phase change in the tree Eucalyptus globulus, which undergoes a dramatic change in vegetative morphology during the juvenile-to-adult transition. Quantitative trait loci analysis in an outcross F2 family derived from crosses between individuals from a coastal population of E. globulus with precocious vegetative phase change and individuals from populations in which vegetative phase change occurs several years later implicated the microRNA EglMIR156.5 as a potential contributor to this heterochronic difference. Additional evidence for the involvement of EglMIR156.5 was provided by its differential expression in trees with early and late phase change. Our findings suggest that changes in the expression of miR156 underlie natural variation in vegetative phase change in E. globulus, and may also explain interspecific differences in the timing of this developmental transition.

Novel Distances for Dollo Data

We investigate distances on binary (presence/absence) data in the context of a Dollo process, where a trait can only arise once on a phylogenetic tree but may be lost many times. We introduce a novel distance, the Additive Dollo Distance (ADD), that applies to data generated under a Dollo model and show that it has some useful theoretical properties including an intriguing link to the LogDet/paralinear distance. Simulations of Dollo data are used to compare a number of binary distances including ADD, LogDet, a restriction-site-based distance, and some simple, but to our knowledge previously unstudied, variations on common binary distances. The simulations suggest that ADD outperforms other distances on Dollo data. Interestingly, we found that the LogDet distance performs poorly in the context of a Dollo process; this may have implications for its use in connection with conditioned genome reconstruction. We apply the ADD to two Diversity Arrays Technology data sets, one that broadly covers Eucalyptus species and one that focuses on the Eucalyptus series Adnataria. We also reanalyze gene family presence/absence data from bacterial genomes obtained from the COG database and compare the results with previous phylogenies estimated using the conditioned genome reconstruction approach. The results for these case studies are largely congruent with previous studies, in some cases giving more phylogenetic resolution.

Genetic differentiation among Australian and Southern Ocean populations of the ubiquitous coccolithophore Emiliania huxleyi (Haptophyta)

Cook S.S., Jones R.C., Vaillancourt R.E. and Hallegraeff G.M. 2013. Genetic differentiation among Australian and Southern Ocean populations of the ubiquitous coccolithophore Emiliania huxleyi (Haptophyta). Phycologia 52: 368–374. DOI: 10.2216/12–111.1 Phytoplankton species with a cosmopolitan distribution are traditionally expected to show little genetic differentiation given the lack of geographical barriers in the ocean. To gain an understanding of the genetic variability within southern hemisphere populations of the coccolithophore Emiliania huxleyi, we used eight microsatellite markers to conduct a population genetic analysis on 273 clonal cultures collected from 11 sites in five ocean current systems south of Australia (Leeuwin, Zeehan, East Australian Currents) and in the Southern Ocean, including within the Antarctic Polar Front. Two of the five currently recognised morphotypes were represented, E. huxleyi var. huxleyi and E. huxleyi var. aurorae. Clonality was absent within sampled populations, suggesting the importance of sexual reproduction in the life cycle of this coccolithophore. Significant genetic differentiation (pairwise population FST range = 0.01–0.09) was apparent among E. huxleyi var. huxleyi populations, much higher than for other cosmopolitan plankton species; thus, gene flow between populations must be low. There was evidence of marked differentiation between Southern Ocean populations of E. huxleyi var. aurorae and E. huxleyi var. huxleyi (pairwise FST range = 0.12–0.16), suggesting the existence of a reproductive, environmental and/or biogeographical barrier between these two varieties.

Expression of a FLOWERING LOCUS T homologue is temporally associated with annual flower bud initiation in Eucalyptus globulus subsp. globulus (Myrtaceae)

The transition to flowering in plants is the result of the balance of endogenous processes and environmental signals that act through a complex genetic pathway that has been studied extensively in annual plants such as Arabidopsis. Perennial trees are characterised by a juvenile non-flowering phase lasting several years followed by an adult phase in which there is repeated cycling between vegetative and reproductive growth. The genetic control of flowering time is potentially more complex in perennials than in annuals and is less understood. Here, we examine the control of flowering in Eucalyptus globulus subsp. globulus, an important forestry species in temperate parts of the world. The E. globulus subsp. globulus homologues of two important flowering genes FLOWERING LOCUS T (FT) and LEAFY (LFY) were isolated and quantitative RT-PCR was used to measure their expression over a 2-year period. The expression of the homologue of FT in E. globulus subsp. globulus leaves was associated with the annual transition from vegetative to reproductive growth (i.e. flower bud initiation). Expression of the LFY homologue was associated with early flower bud development. In a comparison of FT and LFY expression patterns in two clones each of an early and late anthesis genotype, no association between the expression of these genes and the timing of anthesis was shown. Taken together, this indicates that FT and LFY could form part of the flower initiation pathway in Eucalyptus but do not regulate the observed differences in anthesis time.

Genetic differentiation in spite of high gene flow in the dominant rainforest tree of southeastern Australia, Nothofagus cunninghamii.

Nothofagus cunninghamii is a long-lived, wind-pollinated tree species that dominates the cool temperate rainforests of southeastern Australia. The species’ distribution is more or less continuous in western Tasmania but is fragmented elsewhere. However, it is unknown whether this fragmentation has affected the species’ genetic architecture. Thus, we examined N. cunninghamii using 12 nuclear microsatellites and 633 individuals from 18 populations spanning the species’ natural range. Typical of wind-pollinated trees, there was low range-wide genetic structure (FST=0.04) consistent with significant gene flow across most of the species’ range. However, gene flow was not high enough to overcome the effects of drift across some disjunctions. Victorian populations (separated from Tasmania by the 240 km wide Bass Strait) formed a genetic group distinct from Tasmanian populations, had lower diversity (mean allelic richness (Ar)=5.4 in Victoria versus 6.9 in Tasmania) and were significantly more differentiated from one another than those in Tasmania (FST=0.045 in Victoria versus 0.012 in Tasmania). Evidence for bottlenecking was found in small populations that were at least 20 km from other populations. Interestingly, we found little divergence in microsatellite markers between the extremes of genetically based morphological and physiological altitudinal clines suggesting adaptive differentiation is strongly driven by selection because it is likely to be occurring in the presence of gene flow. Even though the cool temperate rainforests of Australia are highly relictual, the species is relatively robust to population fragmentation due to high levels of genetic diversity and gene flow, especially in Tasmania.