Chemical diversity of kanuka: Inter- and intraspecific variation of foliage terpenes and flavanones of Kunzea (Myrtaceae) in Aotearoa/New Zealand.

Kunzea (Myrtaceae) trees and shrubs, generally called kanuka, grow across most of Aotearoa/New Zealand (NZ). With the exception of K. sinclairii, an offshore island endemic, kanuka had been treated as an Australasian species K. ericoides. However, a 2014 taxonomic revision recognized ten species, all endemic to NZ. Kanuka chemistry is less studied than that of its closest relative in NZ, manuka (Leptospermum scoparium), which shows very distinct regional foliage chemotypes. We have used a miniaturized method with GC and 1H NMR to analyze foliage chemistry of voucher specimens from across the geographic ranges of the ten NZ Kunzea species. We found common mono- and sesquiterpenes, with α-pinene dominant in all samples, but only traces of antimicrobial triketones. Two unusual flavanones, with unsubstituted B-rings and known bioactivity against Phytophthora, did distinguish some of the samples. 5,7-Dihydroxy-6,8-dimethyl flavanone was only found at high concentrations in the three K. sinclairii samples in this study's sample set, but this compound has separately been reported in K. robusta samples from a nearby region. Therefore none of the NZ Kunzea species was distinguished by the chemistry analyzed in this study, but there is a possibility of regional flavonoid chemotypes cutting across the species boundaries.

Genetic structuring of the coastal herb Arthropodium cirratum (Asparagaceae) is shaped by low gene flow, hybridization and prehistoric translocation.

We examined the genetic structuring of rengarenga (Arthropodium cirratum; Asparagaceae), an endemic New Zealand coastal herb, using nuclear microsatellite markers. This species was brought into cultivation by Maori within the last 700-800 years for its edible roots and was transplanted beyond its natural distribution as part of its cultivation. We found very high levels of genetic structuring in the natural populations (FST = 0.84), indicating low levels of gene flow. Reduced genetic diversity was found in the translocated populations, suggesting a large loss of genetic diversity early in the domestication process. The data indicates that rengarenga was brought into cultivation independently at least three times, with the sources of these introductions located within a narrow area encompassing about 250km of coastline. Hybridization was inferred between A. cirratum and the closely related A. bifurcatum, despite A. birfucatum not occurring in the vicinity.

Anemonastrum tenuicaule and A. antucense (Ranunculaceae), new combinations for a New Zealand endemic species and its South American relative.

A rational taxonomic circumscription of genera in tribe Anemoneae (Ranunculaceae) is briefly discussed. It is concluded that, in view of the morphological diversity of the group and recent molecular phylogenetic findings, a moderately narrow approach to the re-circumscription of genera earlier included in Anemone sensu lato is preferable, in particular, with the recognition of the lineage with the base chromosome number x = 7 (Anemone subgen. Anemonidium) as two genera, Hepatica sensu stricto and Anemonastrum in an expanded circumscription (including Anemonidium, Arsenjevia, Jurtsevia, and Tamuria). Following these conclusions, new nomenclatural combinations are proposed for two related species endemic to New Zealand and South America, respectively: Anemonastrum tenuicaule (= Anemone tenuicaulis, Ranunculus tenuicaulis) and Anemonastrum antucense (= Anemone antucensis). Information on typification is updated: the lectotype of Anemone antucensis is the specimen from P and not a specimen from G, and the lectotype of Ranunculus tenuicaulis is a specimen from AK. Biogeographic scenarios already proposed to explain the relationship of these two species and some other South America - New Zealand distribution patterns are discussed. It is concluded that the long-distance dispersal scenario fits best the available data for Anemonastrum. Two host-specific and geographically restricted species of Urosystis parasitizing A. tenuicaule and A. antucense are briefly discussed.

Isolation and characterization of microsatellite loci from Arthropodium cirratum (Asparagaceae).

PREMISE OF THE STUDY: Microsatellite markers were developed for Arthropodium cirratum (Asparagaceae) to study population genetic structure and translocation of this species. These markers were tested for cross-amplification in two other Arthropodium species. METHODS AND RESULTS: Sixteen microsatellite markers were developed from a genomic library and tested in three populations of A. cirratum. The loci exhibited one to five alleles per locus, with private alleles present in each of the populations. Cross-amplification tests in the two other New Zealand Arthropodium species revealed that many of the loci amplify and demonstrate polymorphism in A. bifurcatum. CONCLUSIONS: These markers will be useful for determining genetic structure in A. cirratum and for determining the origins of translocated populations of this species.

Evidence of a Strong Domestication Bottleneck in the Recently Cultivated New Zealand Endemic Root Crop, Arthropodium cirratum (Asparagaceae).

We use chloroplast DNA sequencing to examine aspects of the pre-European Maori cultivation of an endemic New Zealand root crop, Arthropodium cirratum (rengarenga). Researching the early stages of domestication is not possible for the majority of crops, because their cultivation began many thousands of years ago and/or they have been substantially altered by modern breeding methods. We found high levels of genetic variation and structuring characterised the natural distribution of A. cirratum, while the translocated populations only retained low levels of this diversity, indicating a strong bottleneck even at the early stages of this species' cultivation. The high structuring detected at four chloroplast loci within the natural A. cirratum range enabled the putative source(s) of the translocated populations to be identified as most likely located in the eastern Bay of Plenty/East Cape region. The high structuring within A. cirratum also has implications for the conservation of genetic diversity within this species, which has undergone recent declines in both its natural and translocated ranges.

Systematics of Simplicia Kirk (Poaceae, Agrostidinae) - an endemic, threatened New Zealand grass genus.

A new species of the New Zealand endemic grass Simplicia, Simplicia felix is described. The new species is segregated from and compared with Simplicia buchananii and Simplicia laxa. Simplicia felix occurs mostly in lightly shaded areas of seasonally dry alluvial forest. A distribution map and an assessment of the conservation status of the new species are presented. Genetic variation in the genus was examined, building on previously published work but including additional sampling. Analysis of nrDNA ITS and ETS and plastid trnL intron and trnL-F intergenic spacer sequences show Simplicia felix to be more closely related to Simplicia laxa than to Simplicia buchananii. NeighborNet analyses of AFLP profiles for the three species of Simplicia show each to consist of distinct clusters of genotypes well separated from each other.

A revision of the New Zealand Kunzeaericoides (Myrtaceae) complex.

A revision of the New Zealand Kunzeaericoides complex is presented. This paper is the final of a series that has explored the systematics of the New Zealand Kunzea complex using cytological and molecular variation, as well as experimental hybridisations between postulated segregates. As a result of those studies ten species, all endemic to New Zealand, are recognised; seven of these are new. One species, Kunzeatriregensis sp. nov., is endemic to the Three Kings Islands and another species Kunzeasinclairii, endemic to Aotea (Great Barrier Island). The North Island of New Zealand has seven species, Kunzeaamathicola sp. nov., Kunzeasalterae sp. nov., Kunzeaserotina sp. nov., Kunzearobusta sp. nov., Kunzeatenuicaulis sp. nov., Kunzeatoelkenii sp. nov., and Kunzealinearis comb. nov. Of these, Kunzealinearis, Kunzeasalterae, Kunzeatenuicaulis and Kunzeatoelkenii are endemic to the North Island, and Kunzeaamathicola, Kunzearobusta and Kunzeaserotina extend to the South Island which also supports one endemic, Kunzeaericoides. Typifications are published for Leptospermumericoides A.Rich., Leptospermumericoidesvar.linearis Kirk, Leptospermumericoidesvar.microflorum G.Simps., Leptospermumericoidesvar.pubescens Kirk, and Leptospermumsinclairii Kirk, names here all referred to Kunzea. The ecology, conservation, extent of natural hybridisation and some aspects of the ethnobotany (vernacular names) of these Kunzea are also discussed.

Species tree phylogeny and character evolution in the genus Centipeda (Asteraceae): evidence from DNA sequences from coding and non-coding loci from the plastid and nuclear genomes.

A species tree phylogeny of the Australian/New Zealand genus Centipeda (Asteraceae) is estimated based on nucleotide sequence data. We analysed sequences of nuclear ribosomal DNA (ETS, ITS) and three plasmid loci (ndhF, psbA-trnH, and trnL-F) using the multi-species coalescent module in BEAST. A total of 129 individuals from all 10 recognised species of Centipeda were sampled throughout the species distribution ranges, including two subspecies. We conclude that the inferred species tree topology largely conform previous assumptions on species relationships. Centipeda racemosa (Snuffweed) is the sister to remaining species, which is also the only consistently perennial representative in the genus. Centipeda pleiocephala (Tall Sneezeweed) and C. nidiformis (Cotton Sneezeweed) constitute a species pair, as does C. borealis and C. minima (Spreading Sneezeweed), all sharing the symplesiomorphic characters of spherical capitulum and convex receptacle with C. racemosa. Another species group comprising C. thespidioides (Desert Sneezeweed), C. cunninghamii (Old man weed, or Common sneeze-weed), C. crateriformis is well-supported but then include the morphologically aberrant C. aotearoana, all sharing the character of having capitula that mature more slowly relative the subtending shoot. Centipeda elatinoides takes on a weakly supported intermediate position between the two mentioned groups, and is difficult to relate to any of the former groups based on morphological characters.

Parallel polyploid speciation: distinct sympatric gene-pools of recurrently derived allo-octoploid Asplenium ferns.

Although polyploidy is widespread, its significance to the generation of biodiversity remains unclear. Many polyploids have been derived recurrently. For a particular polyploid, gene-flow between the products of independent origin is typical where they come into contact. Here, we use AFLP DNA-fingerprinting and chloroplast DNA sequences to demonstrate parallel polyploid speciation within both of the ferns Asplenium cimmeriorum and A. gracillimum. Both of these taxa comprise at least two allopolyploids, recurrently derived from the same progenitor pair. Each of these allopolyploids remain genetically distinguishable even with extensive sympatry, and could therefore be considered distinct species. To our knowledge, parallel speciation on this scale amongst recurrent polyploids has not been previously reported. With their parallel origins, these 'evolutionary replicates' provide an unrivalled opportunity to investigate how the reproductive barriers and ecological differentiation necessary for speciation arise following polyploidy.

Chemotaxonomy of Pseudowintera: sesquiterpene dialdehyde variants are species markers.

Two sesquiterpene dialdehydes, the 1beta-E-coumaroyl-5alpha-hydroxypolygodial plus the known 1beta-E-cinnamoyl-5alpha-hydroxypolygodial, were isolated from the recently described species Pseudowintera insperata. This discovery is a further example of the rare sesquiterpene dialdehyde coumarate/cinnamate combination being found exclusively in the family Winteraceae. Another sesquiterpene dialdehyde, isopaxidal, with the rare rearranged drimane skeleton, was isolated from Pseudowintera axillaris. The sesquiterpene dialdehyde contents of leaves of 25 individual plants of the four Pseudowintera species, all endemic to New Zealand, were measured by HPLC. P. insperata individuals all had high levels (3.0-6.9% of leaf dry wt.) of the coumarate, P. axillaris had high levels (2.2-6.9%) of paxidal, and Pseudowintera colorata from different areas of New Zealand contained varying levels of polygodial (1.4-2.9%) and 9-deoxymuzigadial (0-2.9%). Therefore the sesquiterpene dialdehydes are good species markers.