Igor V. Bartish

Effects of life history traits and sampling strategies on genetic diversity estimates obtained with RAPD markers in plants

Abstract A compilation of studies using RAPD markers for evaluating population differentiation resulted in 78 estimates of AMOVA-derived Φ ST and 31 estimates of Neis G ST , as well as in 41 estimates of Neis within-population diversity. In outcrossing taxa, estimates of between-population diversity were closely correlated with maximum geographic distance between sampled populations. A corresponding association was not found in selfing taxa. These results suggest that RAPD can be a sensitive method for detection of genetic structuring according to the isolation-by-distance model. However, it also means that sampling strategies, as applied in individual studies, can seriously influence the resulting estimates of between-population diversity. Other sampling strategies, like number of plants per population and number of scored polymorphic markers, do not seem to impart any serious artefacts. As previously verified with allozyme data, RAPD markers showed that long-lived, outcrossing, late successional taxa retain most of their genetic variability within populations. By contrast, annual, selfing and/or early successional taxa allocate most of the genetic variability among populations. Estimates for between- and within-population diversity, respectively, proved to be negatively correlated, as previously reported for allozyme data. The only major discrepancy between allozymes and RAPD markers concerns geographic range; within-population diversity was strongly affected by distributional range of the investigated species in the allozyme data but not in the RAPD data. Moreover, RAPD-based values for between-population diversity increased with increasing distributional range whereas the opposite has been reported in a large allozyme data compilation. Contrary to allozymes, RAPD marker-derived within-population diversity is probably therefore not a very good predictor of total species genetic diversity.

Phylogenetic relationships and differentiation among and within populations of Chaenomeles Lindl. (Rosaceae) estimated with RAPDs and isozymes

Abstract RAPD and isozyme analyses based on numerous markers have been used for the first time to investigate patterns of phenetic and genetic differentiation among and within nine wild populations of the genus Chaenomeles represented by the species C. japonica, C. speciosa, C. cathayensis and C. thibetica. Highly significant correlations were found between the two different marker systems for both phenetic distances and gene diversity estimates. In agreement with previous studies on cultivated Chaenomeles material, C. japonica was clearly differentiated from C. speciosa and C. cathayensis. The recently recognised species C. thibetica appeared to be rather closely related to C. cathayensis. Populations of C. japonica and C. speciosa were considerably more diverse than populations of C. cathayensis and C. thibetica. Correspondingly, most of the total variability could be attributed to the within-population differentiation in the case of C. japonica and C. speciosa, and to the between-population differentiation in the case of C. cathayensis. Differences in mating systems among the species can be suggested as a possible explanation of the results. A discordant pattern was found between RAPDs and isozymes in the analyses of population structure within C. japonica. This may be explained by a higher proportion of non-neutral markers for isozymes than for RAPDs. This finding also shows the importance of using multiple molecular marker systems in studies of population structure.

Phylogenetic relationships among New Caledonian Sapotaceae (Ericales): molecular evidence for generic polyphyly and repeated dispersal

The phylogeny of a representative group of genera and species from the Sapotaceae tribe Chrysophylleae, mainly from Australia and New Caledonia, was studied by jackknife analyses of sequences of nuclear ribosomal DNA. The phylogeny conflicts with current opinions on generic delimitation in Sapotaceae. Pouteria and Niemeyera, as presently circumscribed, are both shown to be nonmonophyletic. In contrast, all species currently assigned to these and other segregate genera confined to Australia, New Caledonia, or neighboring islands, form a supported clade. Earlier classifications in which more genera are recognized may better reflect relationships among New Caledonian taxa. Hence, there is need for a revision of generic boundaries in Chrysophylleae, and particularly within the Pouteria complex, including Leptostylis, Niemeyera, Pichonia, Pouteria pro parte (the main part of section Oligotheca), and Pycnandra. Section Oligotheca have been recognized as the separate genus Planchonella, a monophyletic group that needs to be resurrected. Three clades with strong support in our jackknife analysis have one Australian species that is sister to a relatively large group of New Caledonian endemics, suggesting multiple dispersal events between this small and isolated tropical island and Australia. The phylogeny also suggests an interesting case of a relatively recent and rapid radiation of several lineages of Sapotaceae within New Caledonia.

Phylogeny of Hippophae (Elaeagnaceae) Inferred from Parsimony Analysis of Chloroplast DNA and Morphology

Abstract We here report the results from two parsimony analyses of all 15 recognized taxa in Hippophae (Elaeagnaceae), one based on chloroplast DNA (cpDNA), and one based on a combined data set of morphological characters and cpDNA. The genera Elaeagnus and Shepherdia were used as outgroup taxa. In general, the results are congruent with a previous RAPD study, and partly with some of the previous proposed classifications. Monophyly of Hippophae is strongly supported. The most widespread species, H. rhamnoides, is, in spite of low support, most likely monophyletic and distinguished by a single molecular synapomorphy. Due to weak internal support, we refrain from recognizing any sections within the genus. Three taxa, first published as nomina nuda but used by several authors, are here validated and/or described. These are Hippophae neurocarpa subsp. stellatopilosa, H. goniocarpa, and H. litangensis. The latter two were originally suggested to form one species with two subspecies, but they are clearly not monophyletic, a single lineage of evolution, but rather are sister to two different species in the analyses. Together with earlier information from isozymes (unpubl. data) and RAPDs, we believe they are results of two independent hybridisations and we describe them as species. In conjunction, a maternal mode of cpDNA inheritance is suggested. Communicating Editor: Alan T. Whittemore

Inter- and intraspecific genetic variation in Hippophae (Elaeagnaceae) investigated by RAPD markers

Genetic diversity has been investigated by the application of molecular markers in, for the first time, all the taxa recognised in recent treatises of the genusHippophae. RAPD (random amplified polymorphic DNA) analyses were conducted with 9 decamer primers, which together yielded 219 polymorphic markers. We found 16 fixed RAPD markers, i.e. markers that either occurred in all plants of a population or were absent from all plants. Several of these markers were useful for analysis of interspecific relationships, whereas others can be considered as taxon-specific markers. Clustering of taxa and populations in our neighbour-joining based dendrogram was in good agreement with some recently suggested taxonomic treatises ofHippophae. Amount and distribution of genetic variability varied considerably between species. Partitioning of molecular variance withinH. rhamnoides supported earlier findings that a considerable part of the total variance resides among subspecies (59.6%) Within-population variability also differed considerably. Percentage polymorphic RAPD loci and Lynch and Milligan within-population gene diversity estimates showed relatively high values for some species close to the geographic centre of origin in Central Asia, e.g.H. tibetana and the putatively hybridogenousH. goniocarpa. Spatial autocorrelation analyses performed on 12 populations ofH. rhamnoides revealed positive autocorrelation of allele frequencies when geographic distances ranged from 0 to 700 km, and no or negative autocorrelation at higher distances. At distances between 700 and 1900 km, we observed deviations from the expected values with strongly negative autocorrelation of allele frequencies. A corresponding relationship between geographic and genetic distances could not be found when the analysis instead was based on one population from each of 8 species.

Late Quaternary history of Hippophaë rhamnoides L. (Elaeagnaceae) inferred from chalcone synthase intron (Chsi) sequences and chloroplast DNA variation

Fossil pollen records indicate that Hippophaë rhamnoides (Sea Buckthorn) was widespread on late‐ and early postglacial raw soils throughout much of central and northern Europe, but that Early Holocene reforestation restricted populations to northern coastal habitats, or along mountain streams in the Alps, Pyrenees, and Carpathians. We used sequence variation at the nuclear chalcone synthase intron (Chsi), in conjunction with chloroplast DNA–restriction fragment length polymorphism data, to investigate the intraspecific phylogeny, phylogeographic structure, and expansion demographic history of this dioecious and wind‐pollinated shrub at its range‐wide scale in Europe and Asia Minor. Four major Chsi phylogroups of unresolved relationships were identified with estimated divergences ∼172 000 years ago. Large‐scale phylogeographic structures of nuclear and cytoplasmic markers were congruent in identifying (i) southeastern Europe as the most likely source of colonization into central Europe and Scandinavia, and (ii) the area just north of the Alps as a contact zone between populations from the Alps and the east/central European‐Scandinavian lineage. Coalescence‐based analyses (i.e. nested clade analysis and mismatch distributions) of Chsi variation were able to detect at least four major episodes of population growth, all within about the last 40 000 years. In particular, these analyses identified a nearly synchronized timing of population expansions in various parts of the species’ range in central‐eastern Europe/Asia Minor, most likely correlating with the Younger Dryas Stadial (∼13 000–11 600 years ago). It remains to be established whether the phylogeographic history of H. rhamnoides, and particularly its rapid response to the rapid environmental changes of the Younger Dryas cold snap, is unique to the species, or whether it is shared with other cold‐tolerant shrub (or grassland) species known from late‐glacial raw soils in Europe.

Phylogenetically Poor Plant Communities Receive More Alien Species, Which More Easily Coexist with Natives

Alien species can be a major threat to ecological communities, but we do not know why some community types allow the entry of many more alien species than do others. Here, for the first time, we suggest that evolutionary diversity inherent to the constituent species of a community may determine its present receptiveness to alien species. Using recent large databases from observational studies, we find robust evidence that assemblage of plant community types from few phylogenetic lineages (in plots without aliens) corresponds to higher receptiveness to aliens. Establishment of aliens in phylogenetically poor communities corresponds to increased phylogenetic dispersion of recipient communities and to coexistence with rather than replacement of natives. This coexistence between natives and distantly related aliens in recipient communities of low phylogenetic dispersion may reflect patterns of trait assembly. In communities without aliens, low phylogenetic dispersion corresponds to increased dispersion of most traits, and establishment of aliens corresponds to increased trait concentration. We conclude that if quantified across the tree of life, high biodiversity correlates with decreasing receptiveness to aliens. Low phylogenetic biodiversity, in contrast, facilitates coexistence between natives and aliens even if they share similar trait states.

Multi-gene phylogeny of the pantropical subfamily Chrysophylloideae (Sapotaceae): evidence of generic polyphyly and extensive morphological homoplasy

We present a molecular phylogeny of 26 out of the 28 currently accepted genera in the subfamily Chrysophylloideae (Sapotaceae) using parsimony, parsimony jackknifing, and Bayesian inference. A data matrix of 8984 characters was obtained from DNA sequences of seven chloroplast loci, two nuclear loci, indels coded as binary characters, and morphology. Our phylogenetic reconstruction suggests that Chrysophyllum, Pouteria, and Pradosia, as well as some sections within Chrysophyllum and Pouteria, are all polyphyletic. These taxa were previously described largely on the basis of unique combinations of states for a set of morphological characters. Mapping some of these characters onto one of the most parsimonious trees indicates that the symplesiomorphic flower in the subfamily was probably 5‐merous, had stamens inserted in the tube orifice, staminodes, seeds with foliaceous cotyledons, exserted radicle, and endosperm. These characters have subsequently been lost multiple times and cannot be used as synapomorphies to support broad generic concepts. Despite the high degree of homoplasy some well‐defined clades can be described on the basis of alternative character state combinations. Also, many of these well‐supported clades appear to be restricted to particular geographical areas (e.g. all taxa in Australasia form a monophyletic group). Hence, we suggest that the segregate genera Aningeria, Malacantha, and Martiusella may ultimately be resurrected, and probably also Donella and Gambeya, but their circumscriptions are still unclear. One species, Chrysophyllum cuneifolium, may have originated from a hybridization event between continents where the maternal genome (cpDNA) comes from South America and the nuclear genome comes from Africa.

Phylogeny, diagnostic characters and generic limitation of Australasian Chrysophylloideae (Sapotaceae, Ericales): evidence from ITS sequence data and morphology

Current generic limits in Chrysophylloideae (Sapotaceae) from Australia, New Caledonia and the Pacific islands have been shown not to correspond to monophyletic groups. In particular, revisions of generic boundaries are necessary for Pouteria and Niemeyera. We present the first cladistic study of a large representative sample from these areas based on (i) nuclear ribosomal DNA (nrDNA) sequence data, and (ii) combined data of nrDNA and morphology. The data were analyzed with parsimony jackknifing using equal weights and gaps coded as binary characters. Our results from the two data sets are highly congruent and morphological data often increase support as well as tree resolution. A basal polytomy prevents hypotheses of intergeneric relationships, but several groups receive strong support, and hence, four segregates of Pouteria (Beccariella, Planchonella, Sersalisia and Van‐royena) are resurrected. Four others, Albertisiella, Bureavella, Iteiluma and Pyriluma are rejected. Niemeyera is redefined as a small genus confined to Australia. Generic limits within the sister group to Niemeyera are still unclear, a group that includes Leptostylis and Pycnandra. Furthermore, Van‐royena may have originated from an intergeneric hybridization event. Traditionally used and newly identified morphological characters are scrutinized for their diagnostic value. For instance, the position of stamen insertion within the corolla tube is a strong indication of generic relationship. Unique synapomorphies are rare and genera must be distinguished on character state combinations. Following the results, several taxonomic combinations are necessary (Beccariella brownlessiana, B. macrocarpa, B. singuliflora, B. vieillardii, Pichonia daenikeri, Planchonella asterocarpon, P. dothioense, P. myrsinifolia, P. myrsinodendron and P. xylocarpa).

RAPD-based analysis of genetic diversity and selection of lingonberry (Vaccinium vitis-idaea L.) material for ex situ conservation

Random amplified polymorphic DNA markers were used to assess relatedness and genetic diversity for 15 lingonberry (Vaccinium vitis-idaea) populations. Seven primers yielding 59 polymorphic bands were used to analyse 13 populations, representing ssp. vitis-idaea from Sweden, Finland, Norway, Estonia and Russia, and two populations, representing ssp. minus from Japan and Canada. A cluster analysis and a multidimensional scaling analysis (MDS) showed similar phenetic patterns among populations, with a pronounced geographic grouping in most cases. Significant correlations were obtained between geographic and genetic distances for the entire set of populations as well as for the 13 ssp. vitis-idaea populations. Mean within-population diversity was 0.206 when estimated with Lynch and Milligans index, and 0.431 when estimated with Shannons index, which is in agreement with the mixed mating system reported for lingonberry. Within-population variability accounted for 68.6% of the total variance when all populations were included, and for 78.8% when only populations of ssp. vitis-idaea were analysed. Two different approaches were applied to the selection of plant material for a potential gene bank: (1) a hierarchical sampling strategy based on a cluster analysis and (2) the Maximum genetic diversity program, developed for the establishment of core collections. Random sampling was undertaken for comparisons with the selected data sets. The most diverse and representative set of lingonberry specimens was obtained when samples were selected with the Maximum diversity program.