Rafnia (Fabaceae, Crotalarieae) is sub endemic to the fynbos region of South Africa with one species extending to KwaZulu-Natal. Morphological analysis was unable to resolve the relationships within the genus but provided a working hypothesis for future studies. A molecular phylogeny of Rafnia (Fabaceae) is presented. Sequenced data was obtained from five regions of the chloroplast genome (rps16 intron, accD-psa1 spacer, psbA-trnH intergenic spacer, trnL intron and trnL-F intergenic spacer) and one region of the nuclear genome (ITS). Congruence between the plastid and nuclear datasets were very low and combination of datasets was difficult to justify. Several explanations for the incongruent datasets are proposed. In addition to the species-level analyses of Rafnia, a higher-level analysis was carried out with additional taxa sampled from across the ‘core genistoids’. The resulting phylogeny was used to produce an estimate for the age of Rafnia. / Dr. M. van der Bank

Lachnaea L. (Thymelaeaceae), a genus established in 1753, is endemic to the Cape Floristic Region (CFR) of southern Africa. It demonstrates a high level of regional endemism, with 55% of the species endemic to one of the six centres in the CFR. The first detailed taxonomic account of the genus was published by Meisner in 1840 based on the collections of Drège. Three sections were recognised, namely Sphaeroclinium Meisn., Conoclinium Meisn., and Microclinium Meisn., based mainly on the inflorescence structure. The circumscription of Lachnaea has been extended to include all five species previously placed in Cryptadenia Meisn. and at present 40 species and two subspecies are recognised. In this study a detailed species-level phylogenetic tree for Lachnaea was produced using 3 283bp of plastid DNA and 679bp of nuclear DNA for 38 accessions and various outgroups. The loci sequenced were trnL intron and trnL-F spacer, rbcL, rps16 intron and the nuclear gene ITS. Parsimony and Bayesian analyses yielded identical relationships and these are consistant with morphological data, producing the best-supported phylogenetic assesment currently available for the genus. Five informal groupings are also described in the study. The start of the diversification of Lachnaea was estimated at 7.2Mya with a major proliferation around 6Mya. Sympatric speciation in this genus appears to be plausible with patterns suggesting that altitudinal differences and pollinators have been involved in the speciation process. / Dr. M. van der Bank

The tribe Podalyrieae is a diverse and interesting group with most of its species found in the Cape Floristic Region (120 out of 128 species), the smallest floral kingdom and the subject of intense conservation interest. DNA sequence data from nuclear ribosomal internal transcribed spacer, including the 5.8s coding region, were used to reconstruct the phylogeny of this Cape Floral Clade. Results support a transfer of Cadia (Sophoreae) to Podalyrieae. Cyclopia is well imbedded in a paraphyletic Podalyriinae and not sister to the tribe as previously suggested. A broader concept of Podalyrieae, which include Podalyriinae, Xiphothecinae and Cadia is suggested. Dating the nodes of the phylogenetic tree using non-parametric rate smoothing with a fossil calibration point indicates that the root node of Podalyrieae (+ Cadia) is 34.35MYA. / Dr. M. van der Bank

The flora of New Zealand has evolved largely in the absence of terrestrial mammals, the predominant frugivore guilds being birds and reptiles. The evolution of divergent fruit traits in New Zealand may be a consequence of different selection pressure by these two guilds, and two contrasting putative dispersal syndromes have been proposed for New Zealand fleshy fruited plants. Coprosma (Rubiaceae: subtribe Coprosminae) is a speciose and morphologically diverse genus, which is distributed throughout the South Pacific and which also produces variably coloured drupes. It was selected as a model genus to investigate the evolution of fruit traits within this context. For this purpose, a molecular phylogeny for the subtribe Coprosminae and Coprosma was inferred using parsimony, likelihood and split decomposition analysis on sequences from the 16rps intron of cpDNA and ETS and ITS region of nrDNA. Up to 32 species were included in the subtribal analyses, whilst 53 species of Coprosma were used in subgeneric studies. The basis for the variety of fruit colours seen in New Zealand was determined using histology and pigment extractions. To test the assumption that fruit traits among species evolved in concert under selection from frugivore guilds, fruit shape, size and nutrient composition were determined in order to test predictions that these would co-vary with fruit colour. In the Coprosminae, fleshy drupes have arisen from dry fruited schizocarps and with two possible reversals to semi-dry drupes. Within Coprosma, fruit colour was found to be evolutionarily labile and varied both among and within lineages, particularly within two main groups where fruit colour had shifted from orange to blue and white, or red, pink and black colours respectively. The evolution of novel (non-orange) fruit colours was restricted to New Zealand, as was the small-leaved, divaricate growth form, the combination of which has been associated with a putative reptile dispersal syndrome. Several trans-oceanic dispersals out of New Zealand were also inferred from the phylogeny, these extending into Australia, New Guinea and Hawai�i. In these instances, fruit colour did not appear change markedly after establishment in a new country. Within New Zealand, fruit sizes were found to vary with colour as predicted, although the majority of species produced small (< 8 mm), elliptical fruits. There was no clear association between fruit colour and fruit nutrient composition. It would appear that these small, succulent, carbohydrate-rich and lipid-poor fruits cater to a wide range of generalist frugivores. The variety of fruit colours in Coprosma stemmed from qualitative and quantitative differences in carotenoid and anthocyanin composition. The genetic basis for the control of these pathways is currently unknown, but it is assumed that a few regulatory genes can control a substantial amount of phenotypic variation. Considering the evolutionary history of Coprosma, it would appear that a history of recent and rapid speciation, hybridisation and reticulate evolution may have increased the tempo of fruit colour evolution in the genus.

The flora of New Zealand has evolved largely in the absence of terrestrial mammals, the predominant frugivore guilds being birds and reptiles. The evolution of divergent fruit traits in New Zealand may be a consequence of different selection pressure by these two guilds, and two contrasting putative dispersal syndromes have been proposed for New Zealand fleshy fruited plants. Coprosma (Rubiaceae: subtribe Coprosminae) is a speciose and morphologically diverse genus, which is distributed throughout the South Pacific and which also produces variably coloured drupes. It was selected as a model genus to investigate the evolution of fruit traits within this context. For this purpose, a molecular phylogeny for the subtribe Coprosminae and Coprosma was inferred using parsimony, likelihood and split decomposition analysis on sequences from the 16rps intron of cpDNA and ETS and ITS region of nrDNA. Up to 32 species were included in the subtribal analyses, whilst 53 species of Coprosma were used in subgeneric studies. The basis for the variety of fruit colours seen in New Zealand was determined using histology and pigment extractions. To test the assumption that fruit traits among species evolved in concert under selection from frugivore guilds, fruit shape, size and nutrient composition were determined in order to test predictions that these would co-vary with fruit colour. In the Coprosminae, fleshy drupes have arisen from dry fruited schizocarps and with two possible reversals to semi-dry drupes. Within Coprosma, fruit colour was found to be evolutionarily labile and varied both among and within lineages, particularly within two main groups where fruit colour had shifted from orange to blue and white, or red, pink and black colours respectively. The evolution of novel (non-orange) fruit colours was restricted to New Zealand, as was the small-leaved, divaricate growth form, the combination of which has been associated with a putative reptile dispersal syndrome. Several trans-oceanic dispersals out of New Zealand were also inferred from the phylogeny, these extending into Australia, New Guinea and Hawai�i. In these instances, fruit colour did not appear change markedly after establishment in a new country. Within New Zealand, fruit sizes were found to vary with colour as predicted, although the majority of species produced small (< 8 mm), elliptical fruits. There was no clear association between fruit colour and fruit nutrient composition. It would appear that these small, succulent, carbohydrate-rich and lipid-poor fruits cater to a wide range of generalist frugivores. The variety of fruit colours in Coprosma stemmed from qualitative and quantitative differences in carotenoid and anthocyanin composition. The genetic basis for the control of these pathways is currently unknown, but it is assumed that a few regulatory genes can control a substantial amount of phenotypic variation. Considering the evolutionary history of Coprosma, it would appear that a history of recent and rapid speciation, hybridisation and reticulate evolution may have increased the tempo of fruit colour evolution in the genus.

Many angiosperms have undergone some series of polyploidization events over the course of their evolutionary history. In these genomes, especially those resulting from multiple autopolyploidization, it may be relatively easy to recognize all the sets of n homeologous chromosomes, but it is much harder, if not impossible, to partition these chromosomes into n subgenomes, each representing one distinct genomic component of chromosomes making up the original polyploid. Thus, if we wish to infer the polyploidization history of the genome, we could make use of all the gene trees inferred from the genes in one set of homeologous chromosomes to construct a consensus tree, but there is no evident way of combining the trees from the different sets because we have no labelling of the chromosomes that is known to be consistent across these sets. We suggest here that lacking a consistent leaf-labelling, the topological structure of the trees may display sufficient resemblance so that a higher level consensus could be revealing of evolutionary history. This would be especially true of the peripheral structures of the tree, likely representing events that occurred more recently and have thus been less obscured by subsequent evolutionary processes. Here, we present a statistical test to assess whether the subgenomes in a polyploid genome could have been added one at a time. The null hypothesis is that the accumulation of chromosomes follows a stochastic process in which transition from one generation to the next is through randomly choosing an edge, and then subdividing this edge in order to link the new internal vertex to a new external vertex. We analyze the probability distributions of a number of peripheral tree substructures, namely leaf- or terminal-pairs, triples and quadruples, arising from this stochastic process, in terms of some exact recurrences. We propose some conjectures regarding the asymptotic behaviours of these distributions. Applying our analysis to a sugarcane genome, we demonstrate that it is unlikely that the accumulation of subgenomes has occurred one at a time in this genome.

Plants colonised land 460 million years ago and charophytes represent the closest living relatives of land plants. The ability to live on land may depend on the presence of certain cell wall polysaccharides such as xyloglucan, a hemicellulose exclusively found in land plants (Popper and Fry, 2003). The cell walls of charophytes are poorly characterised. The aim of this project was to use biochemical techniques to characterise the cell wall polysaccharides of charophytic algae in relation to early land plant phylogeny. Hydrolysis of Coleochaete scutata and Chara vulgaris cell walls in 2 M trifluoroacetic acid yielded predominantly GalA, Gal, Glc and Man residues and also some Ara, Xyl and traces of Fuc and Rha. In addition, hydrolysis of Chara pectin revealed an abundance of an unusual monosaccharide, 3-O-methyl-D-galactose, which was structurally identified by a series of 1-D and 2D NMR spectroscopy by COSY, TOCSY, NOESY and HSQC. 3-O-Methyl-D-galactose is more commonly found in lycophyte cell walls where its presence has been suggested to be related to lycophytes’ evolutionarily isolated position (Popper et al., 2001). The newly discovered presence of 3-O-methyl-D-galactose in charophyte pectin suggests that this polymer may be more complex than previously thought. Coleochaete and Chara hemicellulose extracts were fractionated by anion-exchange chromatography into five classes. A strongly anionic fraction from Chara hemicellulose was found to be rich in Glc, Xyl, Gal and Fuc suggestive of a xyloglucan-like polysaccharide. However, XEG was unable to produce diagnostic xyloglucan oligosaccharides in either Coleochaete or Chara hemicelluloses. Xylanase and mannanase digestion of Coleochaete and Chara hemicelluloses gave xylan- and mannan-oligosaccharides. Furthermore, lichenase digestion of Coleochaete hemicellulose yielded an unusual octasaccharide composed of approximately equimolar xylose and glucose. My work has shown that charophyte cell walls are a source of undiscovered monosaccharides and potentially novel pectic and hemicellulosic domains which may have important functions in enabling the successful colonisation of land by plants.

At maturity, spermatozoids of the green alga Chara vulgaris are biflagellated, contain little cytoplasm, and coil for approximately 2 1/4 gyres within the mother cell wall. The anterior of the cell contains an ovoid headpiece anchoring two slightly staggered basal bodies that are positioned above and directly in front of approximately 30 linearly arranged mitochondria. An elongated stellate pattern occupies the transition zone between the BBs and axonemes. Flagella emerge from the cell just in front of the nucleus and encircle the full length of the spermatozoid. The spline comprises a maximum of 38 microtubules surrounding the anterior mitochondria and gradually decreases posteriorly to a minimum of 11. The dense nucleus is narrow, cylindrical, and occupies the central revolution of the cell. Six starch-laden plastids and associated mitochondria are linearly arranged at the cell posterior. Phylogenetic analyses of charalean taxa and archegoniates based on spermatogenesis strongly support the order Charales, with Nitella as the sister group to Chara. Diagnostic features of Chara spermatozoids include absence of a lamellar strip and axonemes embedded in the cell for almost the entire length of the anterior mitochondria. Potential relationships among Charales, Coleochaetales and archegoniates are evaluated in regards to the probable course of evolution of streamlined biflagellated gametes.

This study examines the role of deffensive host-plant traits in structuring the community of leaf-chewing insects living on willows. Host-plant phylogeny was reconstructed and leaf morphology and content of three different groups of secondary metabolites were measured. Relationships between defensive leaf traits were examined and their influence on insect community structure was analyzed.