Molecules and morphology in concert: tests of some hypotheses in arctic Potentilla (Rosaceae)

We developed a combined molecular and morphological approach to unravel complex variation at low taxonomic levels, exemplified by some arctic members of Potentilla. Twenty-one populations from Svalbard were analyzed for random amplified polymorphic DNAs (RAPDs) and 64 morphological characters to test the hypotheses that (1) the P. nivea complex (section Niveae) consists of three taxa (P. chamissonis, P. insularis, and P. nivea), (2) three "eco-morphotypes" in P. pulchella (section Multifidae) should be considered different taxa, and (3) P. insularis originated as an intersectional hybrid (Niveae × Multifidae). Twenty-two RAPD multilocus phenotypes were observed in the 136 plants analyzed based on 35 markers. Three fairly distinct groups of RAPD phenotypes were identified in the P. nivea complex based on multivariate analyses and an analysis of molecular variance (AMOVA; 77.6% among-group variation). The variation within the P. nivea complex was more or less continuous in multivariate analyses of the morphological data. We identified, however, several individual morphological characters that separated unambiguously among the three groups of RAPD phenotypes, revealing that these groups correspond to the previously hypothesized taxa. Many identical RAPD multilocus phenotypes were observed in the "eco-morphotypes" of P. pulchella, suggesting that its conspicuous morphological variation is caused by plasticity or by genetic variation at a small number of loci. The hypothesis of the hybrid origin of P. insularis was not supported by the RAPD data. Overall, very little RAPD variation was observed within populations of the four taxa (2.1-16.7% in AMOVA analyses; average genotypic diversity, D, was 0.10-0.30). We conclude that detailed, concerted analysis of molecules and morphology is a powerful tool in low-level taxonomy.     

Genetic divergence, range expansion and possible homoploid hybrid speciation among pine species in Northeast China

Although homoploid hybrid speciation in plants is probably more common than previously realized, there are few well-documented cases of homoploid hybrid origin in conifers. We examined genetic divergence between two currently widespread pines in Northeast China, Pinus sylvestris var. mongolica and Pinus densiflora, and also whether two narrowly distributed pines in the same region, Pinus funebris and Pinus takahasii, might have originated from the two widespread species by homoploid hybrid speciation. Our results, based on population genetic analysis of chloroplast (cp), mitochondrial (mt) DNA, and nuclear gene sequence variation, showed that the two widespread species were divergent for both cp- and mtDNA variation, and also for haplotype variation at two of eight nuclear gene loci surveyed. Our analysis further indicated that P. sylvestris var. mongolica and P. densiflora remained allopatric during the most severe Quaternary glacial period that occurred in Northeast China, but subsequently exhibited rapid range expansions. P. funebris and P. takahasii, were found to contain a mixture of chlorotypes and nuclear haplotypes that distinguish P. sylvestris var. mongolica and P. densiflora, in support of the hypothesis that they possibly originated via homoploid hybrid speciation following secondary contact and hybridization between P. sylvestris var. mongolica and P. densiflora.     

Patterns of allozyme and morphological differentiation in the Carex flava complex (Cyperaceae) in Fennoscandiaxs

Allozyme and morphological differentiation patterns within the Carex flava complex in Fennoscandia were described by means of multivariate analyses. As a result of these studies, and considering also data on ecology, distribution and hybridization, the following taxa were recognized: C. flava with no infraspecific taxa, C. lepidocarpa with two subspecies, ssp. lepidocarpa and ssp. jemtlandica, C. demissa with no infraspecific taxa, and C. oederi with three varieties, var. oederi , var. bergrothii , and var. pulchella. Carex lepidocarpa ssp. jemtlandica was allozymically divergent from ssp. lepidocarpa , indicating a long period of isolation of the two subspecies. A poor morphological separation in some localities is apparently due to secondary contacts in connection with recolonization of Fennoscandia after the last ice age, resulting in the formation of fertile hybrids and back-crosses. The subdivision of C. oederi into three varieties is supported by ecological data and it is suggested that they have originated by parallel ecotype formation under similar selective regimes in different areas. However, the varietal limits are not supported by differentiation in neutral (allozyme) characters. A phylogenetic hypothesis was constructed by means of neighbour-joining based allozyme data and it was indicated that C. demissa may be more closely related to C. lepidocarpa than to C. oederi. Previous treatments that have reduced the C. flava complex to one or two species may have been based on inappropriate species concepts, too coarse methods of analysis of morphological data, ignorance of patterns of co-occurrence and hybridization, or a combination of these factors.     

Long day stimulation of dry matter production in Poa alpina along a latitudinal gradient in Norway

Photoperiod extension with weak include from 8 to 24 h stimulated relative growth rate (RGR) of Poa alpina up to 20%. The response was greatest in young plants at low temperatures. The leaf area ratio (LAR) increased by about 50% which more than compensated for a reduction in net assimilation rate (NAR) by about 20%. The effect of daylength extension on RGR was similar for an arctic ecotype of P. alpina from Svalbard, an alpine ecotype from southern Norway and a borealtemperate ecotype from southern Norway. The main changes in the growth parameters occurred over the 13 to 17 h photoperiod range in P. alpina from both Svalbard and Rondane, although plants at Svalbard are exposed to midnight sun throughout their growing season. The ecotype of P. alpina from southern Norway had fewer tillers and allocated less dry matter to the roots than the two other ecotypes.     

Genetic diversity in populations of plants with different breeding and dispersal strategies in a free-flowing boreal river system

We have studied the genetic diversity of three plant species: Angelica archangelica (Apiacieae), Bistorta vivipara (Polygonaceae) and Viscaria alpina (Caryophyllaceae) along the free-flowing Vindel River in northern Sweden. The plants differ in reproductive strategy. A archangelica and V. alpina are insect pollinated outbreeders while B. vivipara reproduces with apomixis through bulbils. The seeds of A. archangelica may float for over a year, while the propagules (seeds and bulbils, respectively) of V. alpina and B. vivipara float for less than two days. Genetic diversity was assessed using starch gel electrophoresis. The clonal diversity of B. vivipara measured by Simpson's index (D) ranged between 0.78 and 0.99. Only a few clones were shared between localities. The average percentages polymorphic loci and mean He based on polymorphic loci for V. alpina over all localities were 23.1 and 0.15, respectively. Wright's F-statistics showed a significant overall deficit of heterozygotes. The diversity of A. archangelica was found to increase downstream. Genetic diversity of each species is sufficiently high to be used in studies on hydrochory. Dispersal appears to be related to the floating ability of progagules.     

Chromosome counts in the Carex flava complex (Cyperaceae) in Finland

Chromosome numbers from 17 Finnish populations of six taxa in the Carex flava complex were examined. Special attention was given to taxa claimed to be endemic to Fennoscandia. The meiotic counts for C. flava n = 30, C. viridula var. viridula n = 35 and var. pulchella n = 35 are in accordance with previous reports. The counts for C. jemtlandica n = 34, C. bergrothii n = 35, and C. kotilaini n = 34 are new. The hybrid C. flava × kotilaini reveals severely disturbed meiosis with many uni- and multivalents. The results are briefly discussed in relation to morphological data. Carex jemtlandica should evidently be included in C. lepidocarpa. Carex bergrothii and C. kotilaini belong to the polymorphic C. viridula , but the latter probably consitute a specific cytotype.     

Litter of the hemiparasite<Emphasis Type="Italic"> Bartsia alpina</Emphasis> enhances plant growth: evidence for a functional role in nutrient cycling

Hemiparasitic angiosperms concentrate nutrients in their leaves and also produce high quality litter, which can decompose faster and release more nutrients than that of surrounding species. The impact of these litters on plant growth may be particularly important in nutrient-poor communities where hemiparisites can be abundant, such as the sub-Arctic. We tested the hypothesis that plant growth is enhanced by the litter of the hemiparasite Bartsia alpina, in comparison with litter of co-occurring dwarf shrub species, using a pot based bioassay approach. Growth of Betula nana and Poa alpina was up to 51% and 41% greater, respectively, in the presence of Bartsia alpina litter than when grown with dwarf shrub litter (Vaccinium uliginosum, Betula nana and Empetrum nigrum subsp. hermaphroditum). The nutrient concentrations of Betula nana plants grown with Bartsia alpina litter were almost double those of plants grown with dwarf shrub litter, and a significantly greater proportion of biomass was allocated to shoots rather than roots, strongly suggesting that nutrient availability was higher where Bartsia alpina litter was present. The presence of litter from dwarf shrubs, or the moss Hylocomium splendens, did not reduce the positive effect of Bartsia alpina litter on plant growth. E. nigrum litter did not appear to affect plant growth substantially differently from litter of other dwarf shrub species, despite earlier reports of its allelopathic action. The enhanced nutrient uptake and growth of plants in the presence of Bartsia alpina (and potentially other hemiparasitic species) litter could have important implications for communities in which it occurs, including enhanced survival of seedlings of co-occurring species and increased resource patchiness.     

Genetic diversity and origin of Potamogeton anguillanus (Potamogetonaceae) in Lake Biwa,Japan

We analyzed the genetic variation in Potamogeton anguillanus Koidz. and its putative parents, P. malaianus Miq. and P. perfoliatus L., at five allozyme loci of four enzymes to test the hypothesis of a hybrid origin for P. anguillanus, collected in Lake Biwa, Japan. Alleles diagnostic for either P. malaianus or P. perfoliatus were present at four loci. Of 13 single locus phenotypes (SLPs) of P. anguillanus, eight were phenotypes that were expected in F(1) hybrids between P. malaianus and P. perfoliatus. Two SLPs were different from those expected in F(1) hybrids but could have resulted from segregation of parental alleles in later generation hybrids. Each of the remaining three SLPs possessed one allele unique to P. anguillanus. Allozyme analyses thus supported the view that P. anguillanus was derived from hybridization between P. malaianus and P. perfoliatus. It seems likely that the genetic diversity of P. anguillanus found previously originated through multiple hybridizations and sexual processes in P. anguillanus. Other processes such as intragenic recombination, mutation, or hybridization with another lineage are also discussed with reference to the origin of unique alleles.     

Conservation genetics and taxonomic status of the rare Kentucky lady's slipper: Cypripedium kentuckiense (Orchidaceae)

Cypripedium kentuckiense is a recently described rare orchid found in Arkansas (predominantly) and in eight other states. Much debate has focused on whether this taxon should be recognized as a distinct species or considered to be an extreme manifestation of the variability present in the widespread taxon Cypripedium parviflorum var. pubescens. In this study, 12 isozyme loci were analyzed for 14 populations of C. parviflorum var. pubescens and eight populations of C. kentuckiense. These data were used to examine the genetic similarity of these taxa, assess whether isozyme data support the continued recognition of C. kentuckiense as a distinct species, and assess whether a newly discovered disjunct Virginia population of C. kentuckiense is genetically isolated from other C. kentuckiense populations. The isozyme data revealed that the two taxa are very closely related with a high interspecific genetic identity. However, C. kentuckiense populations contain a subset of the variation present in C. parviflorum var. pubescens, and they have expected levels of heterozygosity that are one-quarter that of C. parviflorum var. pubescens populations. Cypripedium kentuckiense also possesses one widespread unique allele and a unique multilocus genotype. These data suggest that C. kentuckiense should be recognized as a distinct species, possibly of recent origin from C. parviflorum. Lastly, the isozyme data support the hypothesis that gene flow between the Virginia population and other populations of C. kentuckiense has been restricted.     

Microsatellite diversity of the agriculturally important alpine grass Poa alpina in relation to land use and natural environment

BACKGROUND AND AIMS: The Alpine Meadow Grass Poa alpina is common in subalpine and alpine natural sites and agriculturally used land, where it is an important fodder grass. Natural factors and human land use are supposed to have been shaping its genetic diversity for hundreds of years. The species comprises sexually and vegetatively reproducing plants. The aim of this study was to investigate the effects of agricultural land use, environmental factors and the mode of reproduction on the distribution of its microsatellite diversity within and among populations and to analyse whether its genetic diversity is correlated with plant species diversity in grassland parcels. METHODS: Genetic diversity of P. alpina was assessed with five microsatellite markers for 569 plants originating from 20 natural sites and from 54 grassland parcels of different cultural tradition, land use and altitude in the Swiss Alps. Due to polyploidy and frequent aneuploidy of the species, data analyses were based on the presence of microsatellite bands. KEY RESULTS: A low but significant differentiation was found in microsatellite bands among natural sites and agriculturally used parcels, while their microsatellite band diversity within populations did not differ. An increased differentiation was found in microsatellite bands with increasing geographic distance among parcels, and a differentiation among grazed and mown parcels, and among sexually and vegetatively reproducing populations. Band richness of sampled plants per village was higher for villages where parcels represented more different land-use types. Within populations, microsatellite band diversity was higher in grazed than in mown parcels. CONCLUSIONS: The diversity of human land use in the Alps was associated with genetic diversity of P. alpina. Therefore, the ongoing socio-economically motivated land-use changes, which reduce the number of different land-use types, will affect the genetic diversity of P. alpina negatively.     

Isozyme evidence for the allotriploid origin of Lycoris flavescens (Amaryllidaceae)

 Variation in isozyme patterns from ten populations of the Korean endemic Lycoris species was used to test the hypothesis that L. flavescens originated from natural hybridization between diploid L. chinensis and L. sanguinea var. koreana. Lycoris sanguinea var. koreana shows fixed heterozygosity at four of nine loci assayed, suggesting that this species is an allotetraploid instead of a diploid. Electrophoretic data suggest that Lycoris flavescens is an allotriploid species derived from the hybridization between diploid L. chinensis and tetraploid L. sanguinea var. koreana. The patterns of allelic distribution in populations of L. flavescens suggest multiple origins of the allotriploid. Within the L. flavescens complex, our isozyme data support the recognition of two taxa, L. flavescens and a recently recognized species, L. uydoensis. Received August 28, 2000 Accepted December 27, 2000     

Genetic diversity and mating system of <Emphasis Type="Italic">Pinus brutia</Emphasis> var. <Emphasis Type="Italic">stankewiczii</Emphasis> Sukacz. in small localities of Sudak (Crimea)

The comparative analysis of genetic variation at 12 polymorphic isozyme loci, as well as the mating system, has been carried out in mature trees and their seed progenies in three small localities of Pinus brutia var. stankewiczii Sukacz. in the region between the town of Sudak and settlement of Novyi Svet in Crimea. We found that embryos maintain the same allelic diversity as parent trees; however, their observed heterozygosity, on average, is lower by 37.4%. The significant deviation of genotype distribution from the expected ratios caused by the deficiency of heterozygotes was observed at 8 out of 12 loci. The multilocus estimate of the out-crossing rate (t _m ) in populations varied from 68.9 to 94.9% with the average of 80.7%.     

The origin of cornbelt maize: The isozyme evidence

Historical records show t hat the Midwestern dent corns of the United States originated from hybridization of two landraces, Northern Flint and Southern Dent. We examined the origin of Southern and Midwestern Dents by means of isozyme electrophoresis. Isozyme genotypes were determined for 23 loci in 12 plants each of 32 accessions of Southern Dent. Previously published isozyme data for maize landraces of Mexico and North America and for U.S. Midwestern Dents were included for comparative purposes. The data show that Northern Flint and Southern Dent are among the isozymically most divergent maize landraces. Nei’s genetic identities between populations of these two landraces are very low for conspecific populations (ca. 0.80). Southern Dent of the southeastern U.S. appears closely related to similar dent corns of southern Mexico, supporting a previously published hypothesis that U.S. Southern Dent is largely derived from the dent corns of southern Mexico. The Midwestern Dents, which resulted from crosses of Southern Dent and Northern Flint, are much more like Southern Dent than Northern Flint in their isozyme profile. Similarly, public inbreds show greater affinity to Southern Dent with the exception of sweet corn lines, which resemble Northern Flint in their isozyme allele frequencies. North American public inbreds do not contain appreciable isozymic variation beyond that found in Northern Flint and Southern Dent.     

Population structure in Arabidopsis lyrata: evidence for divergent selection on trichome production

Leaf trichomes may serve several biological functions including protection against herbivores, drought, and UV radiation; and their adaptive value can be expected to vary among environments. The perennial, self-incompatible herb Arabidopsis lyrata is polymorphic for trichome production, and occurs in a glabrous and a trichome-producing form. Controlled crosses indicate that the polymorphism is governed by a single gene, with trichome production being dominant. We examined the hypothesis that trichome production is subject to divergent selection (i.e., directional selection favoring different phenotypes in different populations) by comparing patterns of variation at the locus coding for glabrousness and at eight putatively neutral isozyme loci in Swedish populations of A. lyrata. The genetic diversity (He) and allele number at isozyme loci tended to increase with population size and decreased with latitude of origin, whereas genetic diversity at the locus coding for glabrousness did not vary with population size and increased with latitude of origin. The degree of genetic differentiation at the glabrousness locus was much higher than that at isozyme loci. Genetic identity at isozyme loci was negatively related to geographic distance, suggesting isolation by distance. In contrast, there was no significant correlation between genetic identity at the glabrousness locus and at isozyme loci. The results are consistent with the hypothesis that divergent selection contributes to population differentiation in trichome production in A. lyrata.     

The intergeneric hybrid origin ofPoa labradorica (Poaceae)

Phytogeographic, ecological, morphological, anatomical, cytological, isozyme, and endonuclease restriction site evidence is presented demonstrating thatPoa labradorica is of intergeneric hybrid origin betweenDupontia fisheri subsp.psilosantha (pistillate parent) andPoa eminens (staminate parent).Poa labradorica is endemic to recently deglaciated areas of northern Quebec and Labrador which are within the overlapping range of the putative parents. Incapable of sexual reproduction,P. labradorica dominates large areas of coastal marsh through vigorous production of rhizomes. Most morphological, anatomical, cytological, isozyme, and DNA characters studied inP. labradorica showed states intermediate or additive to those of the putative parents.     

Six actin gene subfamilies map to five chromosomes of Petunia hybrida

The Mitchell variety of Petunia hybrida possesses a superfamily of actin genes which contains between 100 and 200 members that can be divided into at least six highly divergent subfamilies. The segregation of restriction fragment length polymorphisms among 96 plants from two backcrosses between the Violet 23 and Red 51 Petunia varieties and the Violet 23 x Red 51 hybrid was examined using gene-specific probes from six Petunia actin gene subfamilies. These data were compared with the genotypes of each plant at 11 marker loci which are distributed among the seven chromosomes of Petunia and which determine flower, pollen, and isozyme phenotypes. From these analyses, members of these six actin gene subfamilies were mapped to five locations on five Petunia chromosomes: the PAc9, PAc1, PAc4, and PAc2 subfamilies are on chromosomes I, II, III, and VII respectively; the PAc3 and PAc7 subfamilies are tightly linked on chromosome IV. All members of the PAc4 subfamily cosegregated as a cluster of genes. These data are discussed regarding gene amplification in plants.     

Low isozyme diversity in Nordic and central European populations of Vicia pisiformis and V. dumetorum (Fabaceae)

Vicia pisiformis and V. dumetorum are closely related perennial species occurring in small, disjunct populations throughout Europe. In Sweden, they are classified as "vulnerable" and "care-demanding", respectively. They possess low variation in RAPD (Random Amplified Polymorphic DNA) markers but relatively high morphological variation. We estimated the amount and distribution of isozyme diversity within and among marginal (Sweden, Norway) and central (Czech Republic) populations. Vicia dumetorum was monomorphic for all 14 isozyme loci in the 22 populations investigated, whereas V. pisiformis had 2.94 % polymorphic loci and a mean of 1.02 alleles per loci. The differentiation (F_sr) of populations of V pisiformis were 0.848, but no clear geographical pattern was found. Preliminary results from artificial pollination experiments suggests self-compatibility in V. dumetorum . The low overall polymorphism in Swedish and Czech localities in both species is explained in terms of common postglacial colonization events, recent anthropogenic activity, selfing and clonal reproduction. Finally, the implications of the data for conservation genetics is discussed.     

南方红豆杉紫杉烷高含量植株系RAPD初步研究

南方红豆杉（Ｔａｘｕｓｃｈｉｎｅｎｓｉｓｖａｒ． ｍａｉｒｅｉ（Ｌｅｍｅｅ ｅｔＬｅｖｌ．） Ｃｈｅｎｇ ｅｔＬ．Ｋ．Ｆｕ） 是我国提取抗癌药紫杉醇的原料植物之一,但分布于不同地区的南方红豆杉其紫杉醇含量差异很大。应用ＲＡＰＤ技术,用２１ 个引物共扩增２５６ 条条带,其中有２２４ 条有多态性（８７．８ ％） 。结合紫杉烷及紫杉醇含量测定和植物形态分类,初步探讨了紫杉烷高含量植株的分子特征、形态和生境的相互关系。在供试的１２ 个地区２４ 个样品中,提出３ 个地区存在紫杉烷高含量植株系的可能性,为资源利用和保护以及优良品种的培育提供基     

Partitioning of Electrons between the Cytochrome and Alternative Pathways in Intact Roots

To test the hypothesis that the cytochrome pathway is not invariably saturated when the alternative pathway is engaged, we titrated root respiration of several species with KCN (an inhibitor of the cytochrome pathway), both in the absence and presence of an inhibitor of the alternative pathway (salicylhydroxamic acid, SHAM). The slopes of the resultant KCN [rho] plots ([rho]cyt) were then used to determine whether the cytochrome pathway was saturated in each species. The species used were Festuca ovina ssp. ovina L., Phaseolus vulgaris L., and six Poa species (Poa pratensis L., Poa compressa L., Poa trivialis L., Poa alpina L., Poa costiniana Vick., and Poa fawcettiae Vick.). Although the cytochrome pathway was saturated in a number of species (i.e. [rho]cyt values were 1.0), several others exhibited [rho]cyt values of less than 0.5. Alternative pathway capacity correlated negatively with [rho]cyt, with [rho]cyt values of less than 1.0 occurring in tissues in which the alternative pathway capacity was greater than 25 to 30% of total respiration. The species that did not show full engagement of the cytochrome pathway rarely exhibited SHAM inhibition in the absence of KCN. We conclude that this lack of SHAM inhibition is not due to a lack of alternative pathway engagement but rather to the diversion of electrons from the alternative pathway to the unsaturated cytochrome path following the addition of SHAM.     

中国前胡属酯酶同工酶与种类演化和地理分布格局

利用聚丙烯酰胺凝胶电泳方法分析比较我国前胡属(PeucedanumL.)18种4变种的叶片酯酶同工酶酶谱,并结合形态特征和地理分布,从基因位点分布规律出发,探讨该属种间亲缘关系和地理分布格局。前胡属种类的酯酶同工酶酶谱能较好地反映种间的亲缘关系,基因位点随所分析种类的地理分布而呈现出明显的地带性变化。讨论了滨海前胡(P.japonicumThunb.)、马山前胡(P.mashanenseShanetSheh)和广西前胡(P.guangxienseShanetSheh)、石防风〔P.terebinthaceum(Fisch.)Fisch.exTurcz.〕和宽叶石防风〔P.terebinthaceumvar.deltoideam(MakinoexYabe)Makino〕、红前胡(P.rubricauleShanetSheh)和刺尖前胡(P.elegansKomarov),以及华中前胡(P.medicumDunn)和华北前胡(P.harrysmithiiFeddeexWolff)等种类的酯酶同工酶酶谱类型与系统演化的关系。根据前胡属植物区系,将我国前胡属分成7个酯酶同工酶地理表型。