Samples Of Natural Populations Research Articles

Variation in the Floral Morphology of Prosthechea karwinskii (Orchidaceae), a Mexican Endemic Orchid at Risk.

Prosthechea karwinskii is an orchid endemic to Mexico, threatened by the destruction of its habitat and the extraction of specimens to meet its demand for ornamental and religious use. Most of its populations, including the most locally abundant ones, are found in Oaxaca state. Variations in some floral traits have been observed in these populations. We implemented a morphometric analysis to assess their floral variation and identify the most significant characters in the morphological patterns of this orchid. Floral samples were collected from 17 populations of P. karwinskii in Oaxaca, as well as from specimens used as ornaments during Easter in an Oaxacan community (Zaachila), whose origin is unknown. Sampling of natural populations covered the environmental, geographic, and morphological variation of the species. We performed an analysis of variance (ANOVA), principal component analysis (PCA), canonical variate analysis (CVA), and cluster analysis, including 185 individuals and 45 variables (12 of them were discarded in the multivariate analyses due to high correlation). Characters of the column, lateral sepal, and labellum were most informative for the observed morphological patterns. Albarradas showed the greatest morphological differentiation, mainly due to the column. In general, individuals from the same locality tended to overlap more, especially the populations of Jaltianguis and Yahuiche, which were different from the geographically close population of Etla. Teposcolula presented the highest values in perianth characters, unlike Sola_Rancho Viejo. The specimens recovered from religious ornaments were morphologically more similar to those from Yanhuitlan and Etla. This morphometric analysis identified characters as potential taxonomic markers for P. karwinskii and related species, showing its potential to associate specimens of unknown origin with their probable geographical region. Our work encourages working on collaborative conservation strategies to ensure the long-term permanence of both the species and its traditional uses.

Patterns of thermal adaptation in a globally distributed plant pathogen: Local diversity and plasticity reveal two-tier dynamics.

Plant pathogen populations inhabit patchy environments with contrasting, variable thermal conditions. We investigated the diversity of thermal responses in populations sampled over contrasting spatiotemporal scales, to improve our understanding of their dynamics of adaptation to local conditions. Samples of natural populations of the wheat pathogen Zymoseptoria tritici were collected from sites within the Euro‐Mediterranean region subject to a broad range of climatic conditions. We tested for local adaptation, by accounting for the diversity of responses at the individual and population levels on the basis of key thermal performance curve parameters and “thermotype” (groups of individuals with similar thermal responses) composition. The characterization of phenotypic responses and genotypic structure revealed the following: (i) a high degree of individual plasticity and variation in sensitivity to temperature conditions across spatiotemporal scales and populations; and (ii) geographic variation in thermal response among populations, with major alterations due to seasonal patterns over the wheat‐growing season. The seasonal shifts in functional composition suggest that populations are locally structured by selection, contributing to adaptation patterns. Further studies combining selection experiments and modeling are required to determine how functional group selection drives population dynamics and adaptive potential in response to thermal heterogeneity.

A Population Genomic Assessment of Three Decades of Evolution in a Natural Drosophila Population.

Population genetics seeks to illuminate the forces shaping genetic variation, often based on a single snapshot of genomic variation. However, utilizing multiple sampling times to study changes in allele frequencies can help clarify the relative roles of neutral and non-neutral forces on short time scales. This study compares whole-genome sequence variation of recently collected natural population samples of Drosophila melanogaster against a collection made approximately 35 years prior from the same locality—encompassing roughly 500 generations of evolution. The allele frequency changes between these time points would suggest a relatively small local effective population size on the order of 10,000, significantly smaller than the global effective population size of the species. Some loci display stronger allele frequency changes than would be expected anywhere in the genome under neutrality—most notably the tandem paralogs Cyp6a17 and Cyp6a23, which are impacted by structural variation associated with resistance to pyrethroid insecticides. We find a genome-wide excess of outliers for high genetic differentiation between old and new samples, but a larger number of adaptation targets may have affected SNP-level differentiation versus window differentiation. We also find evidence for strengthening latitudinal allele frequency clines: northern-associated alleles have increased in frequency by an average of nearly 2.5% at SNPs previously identified as clinal outliers, but no such pattern is observed at random SNPs. This project underscores the scientific potential of using multiple sampling time points to investigate how evolution operates in natural populations, by quantifying how genetic variation has changed over ecologically relevant timescales.

Expression of internal reproductive barriers in a germplasm bank accession of the wild potato Solanum chacoense Bitter in three ex situ regeneration cycles

Wild potatoes (Solanum sp.) are important reservoirs of desirable genes for potato breeding. Most of them are diploid and obligate outcrossers, with both sexual and asexual reproduction. Samples of natural populations are ex situ conserved as accessions in germplasm banks for basic and applied ends, and are regenerated when seed number, seed viability or both decrease. The possible action of within accession internal reproductive barriers is not considered in the current seed regeneration protocols, in which 20 to 25 plants are cultivated for the crossing work. Notwithstanding, the effective number of progenitors (Ne) in every seed regeneration cycle might be reduced in relation to the real number (N) of plants if internal reproductive barriers were acting; this process can eventually lead to gene erosion. To ascertain if Ne = N in seed regeneration of one randomly provided accession of S. chacoense Bitter, pollen viability and size, plus pollen-pistil relationships and number and type (according to endosperm development) of seeds/fruit were studied, respectively, at the individual plant and genotypic combination levels. Two types of barriers, pre-zygotic (pollen-pistil self- and cross-incompatibility) and post-zygotic (male sterility and inviability and weakness of F1 hybrids) were detected in three growing seasons, indicating that not all genotypes in this accession made an equal allele contribution to the next generation. These results should be an alert for germplasm bank curators of wild potatoes and other crop wild relatives. Since the objective of germplasm banks is the ex situ maintenance of the natural genetic diversity, strategies need to be devised based on genetic premises.

The Metabolome of Associations between Xylem-Feeding Insects and their Bacterial Symbionts.

Metabolomics has increasingly led to important insights in chemical ecology by identifying environmentally relevant small molecules that mediate inter-organismal interactions. Nevertheless, the application of metabolomics to investigate interactions between phytophagous insects and their microbial symbionts remains underutilized. Here, we investigated the metabolomes of the bacteriomes (organs bearing symbiotic bacteria) isolated from natural populations of five species of xylem-feeding insects. We identified three patterns. First, the metabolomes varied among the five species, likely influenced by insect phylogeny, food plant and taxonomic identity of the symbionts. Second, the ratio of glutamine: glutamate in the bacteriomes was 0.7-3.6 to 1, indicative of nitrogen-sufficient metabolism and raising the possibility that the insect sustains nitrogen-enriched status of the bacteriomes despite the nitrogen scarcity of the xylem diet. Finally, bacteriomes from insect species bearing genetically-similar symbionts displayed limited variation in their metabolomes, suggesting that the metabolic pattern of the bacteriome metabolic pools is correlated with the genetic repertoire of the symbionts. Altogether, these metabolomic patterns yield specific hypotheses of underlying processes that are testable by wider sampling of natural populations and experimental study.

Divergent demographic patterns and perspectives for conservation of endemic species in extreme environments: a case study of the springsnail Melanopsis etrusca (Gastropoda: Melanopsidae)

AbstractWe studied the distribution, size structure, growth, and diet of the freshwater prosobranch Melanopsis etrusca in experimental and natural population samples. This snail is endemic to isolated headwaters of thermal springs in central Italy. In recent times, several anthropogenic pressures such as water extraction and stream contamination have caused population declines and local extinctions of M. etrusca. Consequently, this species was included on the IUCN Red List of Threatened Species. The streams inhabited by M. etrusca had oligohaline waters; however, some physicochemical variability existed among sites. We found that the two variables most directly related to the patterns of distribution and life history of M. etrusca were water pH and temperature. In particular, individual growth was faster in streams with water temperatures constantly >30°C, which was reflected by a shorter life cycle than in cooler streams (13°–26°C), where snails showed a seasonal pattern of growth. Finally, we observed that individuals in a population that is exposed to extreme temperatures (>35°C) performed a peculiar behavior, spending long periods outside the water. We also found that the diet of M. etrusca reflected the local composition of the periphytic community and was dominated by macrophyte fragments. Our data suggest that heat‐adapted populations of M. etrusca hold the greatest potential for recolonization after collapse events. From a conservation perspective, reintroduction programs could be useful to avoid local extinction or bottleneck effects but at the risk of the loss of evolutionary innovations that may have emerged at the population level.

Genetic diversity and population structure of the African catfish, Clarias gariepinus (Burchell, 1822) in Kenya: implication for conservation and aquaculture

African catfish, Clarias gariepinus, is an important species in aquaculture and fisheries in Kenya. Mitochondrial D-loop control region was used to determine genetic variation and population structure in samples of C. gariepinus from 10 sites including five natural populations (Lakes Victoria (LVG), Kanyaboli (LKG), Turkana (LTA), Baringo (LBA) and Jipe (LJP), and five farms (Sangoro Aquaculture Center (SAN), Sagana Aquaculture Centre (SAG), University of Eldoret Fish Farm (UoE), Kibos Fish Farm (KIB), and Wakhungu Fish Farm (WKU)) in Kenya. Similarly, samples from eight localities (four natural populations: LVG/LKG, LTA, LBA, and four farmed: SAN, SAG, KIB, UoE) were genotyped using six microsatellite DNA loci. For the D-loop control region, samples from natural sites exhibited higher numbers of haplotypes and haplotype diversities compared to farmed samples, and 88.2% of haplotypes were private. All except LJP and LTA shared haplotypes, and the highest number of shared haplotypes (8) was detected in KIB. The 68 haplotypes we found in 268 individuals grouped into five phylogenetic clades: LVG/LKG, LTA, LBA, LJP and SAG. Haplotypes of farmed C. gariepinus mostly have haplotypes typical of LVG/LKG, and some shared haplotypes of the LBA population. Microsatellite analysis showed farmed samples have higher numbers of alleles than natural samples, but higher observed and expected heterozygosity levels were found in samples of natural populations. Fifteen pair-wise comparisons had significantly different FST values. All samples were in Hardy-Weinberg equilibrium. Samples from the eight localities grouped into four genetic clusters (LVG/LKG, LTA, LBA and SAG), indicating genetically distinct populations, which should be considered for aquaculture and conservation.

Evaluation of noninvasive genetic methods for Nubian ibex

Noninvasive genetic methods enable the sampling of natural populations while minimizing detrimental effects on them. However, noninvasive methods are marred by DNA extraction and amplification difficulties that can be mitigated by improved methodology. Past studies have shown that noninvasive genetic protocols are system specific and highlighted the importance of pilot studies in the establishment of genetic monitoring programs. We tested, using a factorial design experiment, the effect of different methods for the collection-preservation and extraction on the amplification of DNA from Nubian ibex (Capra nubiana) fecal samples. We found collection-preservation using paper bags and extraction with QIAamp® fast DNA stool mini kit to significantly enhance success rate compared to collection-preservation in ethanol and extraction with QIAamp® DNA mini kit. Our results will contribute to the studies of natural populations of the vulnerable C. nubiana.

Sex identification of the extant mega mammal, the lowland tapir, Tapirus terrestris (Tapiridae, Mammalia), by means of molecular markers: new outlook for non-invasive samples

The knowledge of the sex ratio of threatened populations or species is pivotal since sub-optimal sex ratios can affect negatively the population growth and resilience. The vulnerable species, Tapirus terrestris, is rarely studied through traditional field methods and non-invasive genetic analyses have been employed using feces as source of DNA. Here, we evaluated two molecular markers for sex identification of tapirs. Amelogenin gene (AMEL) which is shorter in the Y chromosome (−20pb), failed in the determination of the sex of tapirs, even after the design of specific primers for the species. SRY marker, co-amplified with zinc-finger gene (ZF), had successful produced the sex identity for blood and scat samples. From a total of 45 feaces, 29 had the sex identified after three PCRS (64.4 %). In this way, these molecular markers are reliable tools for sex ratio determination using non-invasive samples of natural populations of this tapirs.

Towards a molecular taxonomic key of the Aurantioideae subfamily using chloroplastic SNP diagnostic markers of the main clades genotyped by competitive allele-specific PCR.

BackgroundChloroplast DNA is a primary source of molecular variations for phylogenetic analysis of photosynthetic eukaryotes. However, the sequencing and analysis of multiple chloroplastic regions is difficult to apply to large collections or large samples of natural populations. The objective of our work was to demonstrate that a molecular taxonomic key based on easy, scalable and low-cost genotyping method should be developed from a set of Single Nucleotide Polymorphisms (SNPs) diagnostic of well-established clades. It was applied to the Aurantioideae subfamily, the largest group of the Rutaceae family that includes the cultivated citrus species.ResultsThe publicly available nucleotide sequences of eight plastid genomic regions were compared for 79 accessions of the Aurantioideae subfamily to search for SNPs revealing taxonomic differentiation at the inter-tribe, inter-subtribe, inter-genus and interspecific levels. Diagnostic SNPs (DSNPs) were found for 46 of the 54 clade levels analysed. Forty DSNPs were selected to develop KASPar markers and their taxonomic value was tested by genotyping 108 accessions of the Aurantioideae subfamily. Twenty-seven markers diagnostic of 24 clades were validated and they displayed a very high rate of transferability in the Aurantioideae subfamily (only 1.2 % of missing data on average). The UPGMA from the validated markers produced a cladistic organisation that was highly coherent with the previous phylogenetic analysis based on the sequence data of the eight plasmid regions. In particular, the monophyletic origin of the “true citrus” genera plus Oxanthera was validated. However, some clarification remains necessary regarding the organisation of the other wild species of the Citreae tribe.ConclusionsWe validated the concept that with well-established clades, DSNPs can be selected and efficiently transformed into competitive allele-specific PCR markers (KASPar method) allowing cost-effective highly efficient cladistic analysis in large collections at subfamily level. The robustness of this genotyping method is an additional decisive advantage for network collaborative research. The availability of WGS data for the main “true citrus” species should soon make it possible to develop a set of DSNP markers allowing very fine resolution of this very important horticultural group.Electronic supplementary materialThe online version of this article (doi:10.1186/s12863-016-0426-x) contains supplementary material, which is available to authorized users.

Genetic variation of oregano (<i>Origanum vulgare</i> L.) for etheric oil in Albania

In this study are presented the results of studies for oregano in 16 locations (sites) of Albania for the content of the quantity of oils and their fractions.Albaniais the one of the important diversity centers for Oregano because of their adaptation to diverse agro ecological conditions as a result of natural selection. The design of the experiment was based on split plot methods. From the total samples collected, were selected 16 mostly widespread samples of natural populations of origano (Origanum vulgare L. sp. vulgare and sp. hirtum). Essential oil isolation from oregano was performed by hydro distillation. Identification of the components was made by comparing mass spectra of components in essential oils with those from Nist, Wiley and Adams mass spectra libraries. The results show that the total oil content varies from 3.45% to 0.1%. The populations of Oregano in the part of southern Albania have a higher amount of etheric oil compared with those of the North part. The higher content of carvacrol in 55.63%, of the samples was determined in sites of the southern part of Albania, while in the other two investigated area central and north part it was 55.63% and 48.49% respectively. The same indicators were realised also for content of linalool and thymol, while North area has sites with high content of caryophyllen-oxide and β-pinen 3. But, in all sites, the oils of thymol and carvacrolit were dominant.

Relevance of the genetic structure of natural populations, and sampling and classification approaches for conservation and use of wild crop relatives: potato as an example

Crop wild relatives (CWRs) are wild taxa with close genetic relationships to species with direct socioeconomic importance. As essential components of natural habitats and agricultural systems, their conservation and sustainable use are vital. CWRs are ex situ conserved in germplasm banks as samples of natural populations (accessions). Most accessions have been assigned specific status according to the Taxonomic Species Concept, which presupposes that living organisms are at the end of speciation. Thus, the morphological and genetic variability that could be encountered in natural populations is disregarded, negatively affecting conservation of allelic frequencies and effective use in breeding. Passport information usually contains collection date and geographical data, but not reproductive behavior of the sampled population (which defines its genetic structure) and sampling strategies. For ex situ multiplications of original collections, no information is provided on the effectively used strategies. As information on how accessions were composed is lacking, conservation of the allelic frequencies from the originally sampled populations is unknown. Knowledge of reproductive biology of plant populations is of utmost importance to understanding their natural morphological and molecular variability and to developing appropriate methodological approaches for sampling, classifying, and ex situ multiplying to conserve gene frequencies for basic and applied purposes.

Life table of Hypothenemus hampei (Ferrari) in relation to coffee berry phenology under Colombian field conditions

The overlap of generations of coffee berry borer (CBB), Hypothenemus hampei (Ferrari) under field conditions in countries like Colombia hinders the construction of life tables by the sampling of natural populations. In this paper, a field methodology to carry out regular measurements of CBB cohorts inside coffee berries of different ages until harvest, both in coffee trees and in infested berries placed on the ground, is developed and used to compare the life history parameters of CBB. Populations with berries at six ages in three experimental stations (without CBB control) and in a commercial farm in Colombia (with chemical CBB control regularly carried out) were used. The duration of the pre-oviposition period as well as the mortality and survival rates of founder females and the proportion of founders leaving infested berries were strongly influenced by the consistency of berries, with optimum conditions for CBB reproduction as from 120-150 days after flowering. No differences were found between stations for the number of CBB developmental stages; but they had larger values than the commercial farm. The latter also had more than twice the average rate of founders leaving infested berries recorded in the stations. Survival functions (cumulative probabilities of survival) for the pest differed among treatments and between the plant and ground micro-environments. Age of berries at infestation was positively related to the intrinsic rate of increase of borer population; whilst generation time and doubling time were inversely related. No differences were found between sites for the main demographic parameters of the pest.

Haplotype Structure and Divergence at Human and Chimpanzee Serotonin Transporter and Receptor Genes: Implications for Behavioral Disorder Association Analyses

Genetic variation in the human serotonin system has long-been studied because of its functional consequences and links to various behavior-related disorders and it being routinely targeted in research and development for drug therapy. However, aside from clinical studies, little is known about this genetic diversity and how it differs within and between human populations with respect to haplotype structure, which can greatly impact phenotype association studies. In addition, no evolutionary approach among humans and other primates has examined how long- and short-term selective pressures explain existing serotonin variation. Here, we examine DNA sequence variation in natural population samples of 192 human and 40 chimpanzee chromosome sequences for the most commonly implicated approximately 38-kb serotonin transporter (SLC6A4) and approximately 63-kb serotonin 2A receptor (HTR2A) genes. Our comparative population genetic analyses find significant linkage disequilibrium associated with functionally relevant variants in humans, as well as geographic variation for these haplotypes, at both loci. In addition, although amino acid divergence is consistent with purifying selection, promoter and untranslated regions exhibit significantly high divergence in both species lineages. These evolutionary analyses imply that the serotonin system may have accumulated significant regulatory variation over both recent and ancient periods of time in both humans and chimpanzees. We discuss the implications of this variation for disease association studies and for the evolution of behavior-related phenotypes during the divergence of humans and our closest primate relatives.

Hakuna Nematoda: genetic and phenotypic diversity in African isolates of Caenorhabditis elegans and C. briggsae

Caenorhabditis elegans and C. briggsae have many parallels in terms of morphology, life history and breeding system. Both species also share similar low levels of molecular diversity, although the global sampling of natural populations has been limited and geographically biased. In this study, we describe the first cultured isolates of C. elegans and C. briggsae from sub-Saharan Africa. We characterize these samples for patterns of nucleotide polymorphism and vulva precursor cell lineage, and conduct a series of hybrid crosses in C. briggsae to test for genetic incompatibilities. The distribution of genetic diversity confirms a lack of geographic structure to C. elegans sequences but shows genetic differentiation of C. briggsae into three distinct clades that may correspond to three latitudinal ranges. Despite low levels of molecular diversity, we find considerable variation in cell division frequency in African C. elegans for the P3.p vulva precursor cell, and in African C. briggsae for P4.p, a variation that was not previously observed in this species. Hybrid crosses did not reveal major incompatibilities between C. briggsae strains from Africa and elsewhere, and there was some evidence of inbreeding depression. These new African isolates suggest that important ecological factors may be shaping the patterns of diversity in C. briggsae, and that despite many similarities between C. elegans and C. briggsae, there may be more subtle differences in their natural histories than previously appreciated.

Extreme genomic variation in a natural population.

Whole-genome sequence data from samples of natural populations provide fertile grounds for analyses of intraspecific variation and tests of population genetic theory. We show that the urochordate Ciona savignyi, one of the species of ocean-dwelling broadcast spawners commonly known as sea squirts, exhibits the highest rates of single-nucleotide and structural polymorphism ever comprehensively quantified in a multicellular organism. We demonstrate that the cause for the extreme heterozygosity is a large effective population size, and, consistent with prediction by the neutral theory, we find evidence of strong purifying selection. These results constitute in-depth insight into the dynamics of highly polymorphic genomes and provide important empirical support of population genetic theory as it pertains to population size, heterozygosity, and natural selection.

The bacterial species definition in the genomic era.

The bacterial species definition, despite its eminent practical significance for identification, diagnosis, quarantine and diversity surveys, remains a very difficult issue to advance. Genomics now offers novel insights into intra-species diversity and the potential for emergence of a more soundly based system. Although we share the excitement, we argue that it is premature for a universal change to the definition because current knowledge is based on too few phylogenetic groups and too few samples of natural populations. Our analysis of five important bacterial groups suggests, however, that more stringent standards for species may be justifiable when a solid understanding of gene content and ecological distinctiveness becomes available. Our analysis also reveals what is actually encompassed in a species according to the current standards, in terms of whole-genome sequence and gene-content diversity, and shows that this does not correspond to coherent clusters for the environmental Burkholderia and Shewanella genera examined. In contrast, the obligatory pathogens, which have a very restricted ecological niche, do exhibit clusters. Therefore, the idea of biologically meaningful clusters of diversity that applies to most eukaryotes may not be universally applicable in the microbial world, or if such clusters exist, they may be found at different levels of distinction.

Esterases no exame da estrutura populacional de Camu-camu (Myrciaria dubia (Kunth) McVaugh-Myrtaceae)

Dois sistemas enzimáticos (esterase e esterase-D), analisados pela técnica de eletroforese em gel de amido, em folhas jovens de plantas cultivadas em terra firme, de sementes provenientes de três amostras de populações naturais de camu-camu, Myrciaria dubia (Kunth) McVaugh-Myrtaceae, procedentes de Iquitos, Boa Vista e Uatumã, revelaram a presença de 6 locos: Est-1, Est-2, Est-3, Est-4, Est-D1 e Est-D2. Dois dos seis locos gênicos examinados no presente estudo (Est-3 e Est-D2) mostraram-se polimórficos, sendo desse modo considerados valiosos no estudo de caracterização da estrutura populacional da espécie. Os padrões de polimorfismo revelados nos locos Est-3 e Est-D2 de camu-camu, são típicos de enzimas monoméricas e diméricas, respectivamente. O loco Est-3 apresentou um grande desbalanço genético dentro e entre as amostras populacionais examinadas, devido ao excessivo número observado de plantas heterozigóticas em relação ao número esperado. O loco Est-D2 apresentou um polimorfismo exclusivo para os alelos Est-D2¹,Est-D2² e Est-D2³, e um bom balanço genético na amostra populacional de Uatumã. Em função disso, dentre os demais locos gênicos aqui investigados, o loco Est-D2 parece ser o mais adequado para identificação e delimitação de prováveis estoques de camu-camu. Portanto, recomenda-se que esse loco esteja presente na lista dos marcadores isoenzimáticos a serem usados em futuras prospecções sobre genética populacional dessa espécie na região amazônica. Dados sobre a distribuição das freqüências alélicas, estimativas das distâncias genéticas, e estimativas de variação genética nos 6 locos de esterases examinados, foram eficazes na demonstração de diferenças genéticas entre as amostras populacionais examinadas da espécie. Os maiores valores de heterozigozidade média (0,1353); proporção de locos polimórficos (0,33) e número médio de alelos por loco (1,33) revelados na amostra populacional de Uatumã, devem-se ao fato dessa amostra ter apresentado um loco polimórfico a mais (Est-D2), em relação as demais amostras investigadas.

Distribution and sequence analysis of a novel Ty3-like element in natural Saccharomyces paradoxus isolates.

Little is known about the transposable elements of species closely related to Saccharomyces cerevisiae. We present a novel transposable element in Saccharomyces paradoxus, a close congener of S. cerevisiae. Sequence analysis of this element, designated Ty3-1p, indicates that it is a homologue of the S. cerevisiae Ty3 element. Ty3-1p shares 82% nucleotide identity with an S. cerevisiae Ty3 element and appears to be structured identically to Ty3, containing two overlapping open reading frames, six retroviral-like domains, a J domain, and flanking sigma-like elements. A sigma element from Ty3-1p is 75% identical to a Ty3 sigma element. There is no evidence of horizontal transfer of Ty3 in Saccharomyces sensu stricto. We assess the distributions of Ty3p and Ty3 element insertions in natural population samples of S. paradoxus and S. cerevisiae. The S. paradoxus population sample exhibits Ty3p insertions present at a variety of sites at low frequency; this suggests that Ty3p elements are active in the sampled population. The S. cerevisiae population sample exhibits a uniform Ty3 hybridization profile in which all element insertions appear to be fixed. We comment on the possible causes of these contrasting observed distributions (GenBank Accession Nos AY198186 and AY198187).

The use of molecular techniques to resolve relationships among traditional weaving cultivars of Phormium

Molecular techniques ‐ sequencing of the nuclear internal transcribed spacer region (ITS), and fingerprinting of amplified fragment length polymorphisms (AFLP) and intersimple sequence repeats (ISSR) ‐ were investigated for their potential to characterise genetic variation in harakeke (Phormium spp., family Hemerocallidaceae) in New Zealand. ITS showed no variation between species of Phormium. However, both AFLP and ISSR identified genome regions that were polymorphic among intraspecific accessions of Phormium obtained from the National New Zealand Flax Collection. When used as a direct sequencing marker, one AFLP fragment distinguished two major groups of nuclear haplotypes. The partitioning of taxa into these groups was supported by phylogenetic analysis of ISSR fingerprinting profiles. Split decomposition, a network‐building technique, was shown to be useful in representing the sorts of relationships present in this study of intraspecific variation. Further sampling of natural populations is required to understand the origins and present‐day distributions of these accessions; however, the present results show the usefulness of these methods for addressing such ethnobotanic questions.