An approach to determining anthocyanin synthesis enzyme gene expression in an evolutionary context: an example from Erica plukenetii.

BACKGROUND AND AIMS: Floral colour in angiosperms can be controlled by variations in the expression of the genes of the anthocyanin pathway. Floral colour shifts influence pollinator specificity. Multiple shifts in floral colour occurred in the diversification of the genus Erica (Ericaceae), from plesiomorphic pink to, for example, red or white flowers. Variation in anthocyanin gene expression and its effects on floral colour in the red-, pink- and white-flowered Erica plukenetii species complex was investigated. METHODS: Next generation sequencing, reverse transcriptase PCR and real-time reverse transcriptase quantitative PCR were used to quantify anthocyanin gene expression. KEY RESULTS: Non-homologous mutations causing loss of expression of single genes were found, indicating that the cause was likely to be mutations in transcription factor binding sites upstream of the 5'-untranslated region of the genes, and this was confirmed by sequencing. CONCLUSIONS: Independent evolution and subsequent loss of expression of anthocyanin genes may have influenced diversification in the E. plukenetii species complex. The approach developed here should find more general application in studies on the role of floral colour shifts in diversification.

The biodiversity hotspot as evolutionary hot-bed: spectacular radiation of Erica in the Cape Floristic Region.

BACKGROUND: The disproportionate species richness of the world's biodiversity hotspots could be explained by low extinction (the evolutionary "museum") and/or high speciation (the "hot-bed") models. We test these models using the largest of the species rich plant groups that characterise the botanically diverse Cape Floristic Region (CFR): the genus Erica L. We generate a novel phylogenetic hypothesis informed by nuclear and plastid DNA sequences of c. 60 % of the c. 800 Erica species (of which 690 are endemic to the CFR), and use this to estimate clade ages (using RELTIME; BEAST), net diversification rates (GEIGER), and shifts in rates of diversification in different areas (BAMM; MuSSE). RESULTS: The diversity of Erica species in the CFR is the result of a single radiation within the last c. 15 million years. Compared to ancestral lineages in the Palearctic, the rate of speciation accelerated across Africa and Madagascar, with a further burst of speciation within the CFR that also exceeds the net diversification rates of other Cape clades. CONCLUSIONS: Erica exemplifies the "hotbed" model of assemblage through recent speciation, implying that with the advent of the modern Cape a multitude of new niches opened and were successively occupied through local species diversification.

Testing reticulate versus coalescent origins of Erica lusitanica using a species phylogeny of the northern heathers (Ericeae, Ericaceae).

Whilst most of the immense species richness of heathers (Calluna, Daboecia and Erica: Ericeae; Ericaceae) is endemic to Africa, particularly the Cape Floristic Region, the oldest lineages are found in the Northern Hemisphere. We present phylogenetic hypotheses for the major clades of Ericeae represented by multiple accessions of all northern Erica species and placeholder taxa for the large nested African/Madagascan clade. We identified consistent, strongly supported conflict between gene trees inferred from ITS and chloroplast DNA sequences with regard to the position of Erica lusitanica. We used coalescent simulations to test whether this conflict could be explained by coalescent stochasticity, as opposed to reticulation (e.g. hybridisation), given estimates of clade ages, generation time and effective population sizes (Ne). A standard approach, comparing overall differences between real and simulated trees, could not clearly reject coalescence. However, additional simulations showed that at the (higher) Ne necessary to explain conflict in E. lusitanica, further topological conflict would also be expected. Ancient hybridisation between ancestors of northern species is therefore a plausible scenario to explain the origin of E. lusitanica, and its morphological similarities to E. arborea. Assuming either process influences the results of species tree and further evolutionary inference. The coalescence scenario is equivocal with regard the standing hypothesis of stepping stone dispersal of Erica from Europe into Africa; whereas reticulate evolution in E. lusitanica would imply that the colonisation of Tropical East Africa by E. arborea instead occurred independently of dispersals within the rest of the African/Madagascan clade.

Historical biogeography of two cosmopolitan families of flowering plants: Annonaceae and Rhamnaceae.

Annonaceae are a pantropically distributed family found predominantly in rainforests, so they are megathermal taxa, whereas Rhamnaceae are a cosmopolitan family that tend to be found in xeric regions and may be classified as mesothermal. Phylogenetic analyses of these families are presented based on rbcL and trnL-F plastid DNA sequences. Likelihood ratio tests revealed rate heterogeneity in both phylogenetic trees and they were therefore made ultrametric using non-parametric rate smoothing and penalized likelihood. Divergence times were then estimated using fossil calibration points. The historical biogeography of these families that are species rich in different biomes is discussed and compared with other published reconstructions. Rhamnaceae and most lineages within Annonaceae are too young to have had their distribution patterns influenced by break-up of previously connected Gondwanan landmasses. Contrasts in the degree of geographical structure between these two families may be explained by differences in age and dispersal capability. In both groups, long-distance dispersal appears to have played a more significant role in establishing modern patterns than had previously been assumed. Both families also contain examples of recent diversification of species-rich lineages. An understanding of the processes responsible for shaping the distribution patterns of these families has contributed to our understanding of the historical assembly of the biomes that they occupy.

An outbreak of strangles in young ponies.

A natural outbreak of strangles occurred in a group of 19 young experimental ponies. The disease was diagnosed in 11 of them within two days of their arrival at Glasgow University veterinary school and five others developed clinical signs within a further four days, a morbidity rate of 84 per cent. All of the affected ponies had typical signs of strangles including dullness, anorexia, pyrexia, regional lymphadenitis, occasionally with rupture of the lymph node, conjunctivitis and a mucopurulent nasal discharge. Nine of the affected ponies were destroyed during the clinical phase of the disease for post mortem studies. The clinical disease in the remaining animals lasted approximately 21 days although one pony had to be destroyed 10 days after the onset of clinical signs because of the development of septic arthritis. All 16 affected animals exhibited peripheral blood neutrophilia and high plasma fibrinogen levels. beta haemolytic streptococci were isolated by nasopharyngeal swabbing from 18 of the 19 ponies. Streptococcus equi was confirmed only in three animals within the first four days of the outbreak. The majority of the other isolates identified to species were S zooepidemicus. beta haemolytic streptococci were still present in six ponies 40 days after they had clinically recovered and were isolated regularly from three ponies which did not develop clinical strangles but remained in contract with affected animals throughout the study.

A scanning electron microscopic study of the equine upper respiratory tract.

The surface features of the upper respiratory tract of 20 clinically normal horses of various ages and types were studied with scanning electron microscopy. In the rostral part of the nasal cavity, there was a wide zone of non-ciliated epithelium whereas, caudally, the surface was well ciliated. This latter type of epithelium extended into the nasopharynx and guttural pouches although scattered areas of non-ciliated microvillous cells were also found.

An ultrastructural study of the equine lower respiratory tract.

The surface features of the lower respiratory tract of 20 clinically normal horses of different ages and types were studied with scanning electron microscopy. Parallel light microscopical and transmission electron microscopical studies were also carried out. The ciliary carpet was virtually complete from the trachea to the lobar bronchi. In small bronchi, ciliation was less complete allowing numerous non-ciliated mucous cells to become obvious. The terminal bronchioles, populated mainly by non-ciliated bronchiolar epithelial cells, had an abrupt junction with alveolar ducts. Interalveolar pores were common particularly in older horses.