Micropropagation of an endangered plant species: Coronopus navasii (Brassicaceae)

Plantlets of Coronopus navasii, an endangered species from SE Spain, were successfully regenerated from shoot and root segments excised from young seedlings. Initiation of multiple buds and development of leaves were obtained in MS modified medium plus l mg/l BAP and 0.1 mg/l NAA. Rooting was achieved by transfer of the isolated shoots to fresh MS medium without plant growth regulators. Plant survival of 47% was obtained six weeks after removal from in vitro culture conditions.Abbreviations BAP 6-benzylaminopurine - NAA naphthaleneacetic acid - MS medium see Murashige and Skoog 1962     

A new Biscutella (Brassicaceae) species from the western Balearic Islands

A new diploid species, Biscutella ebusitana is described from coastal cliffs of the western Balearic islands. It is related to B. sempervirens L. and B. montana Cav. from which it can be distinguished by a combination of vegetative (shape, size, petiole and hairs of leaves, petiolatc leaves) and reproductive features (shape of inflorescence, calyx indument). Earlier Balearic records of B. frutescens Coss. should be referred to B. ebusitana.     

Infraspecific genetic variation in Biscutella laevigata (Brassicaceae): new focus on Irene Manton's hypothesis

 Genetic variation in 42 populations throughout the range of Biscutella laevigata L. (Brassicaceae), a morphologically variable central European species, has been investigated by enzyme electrophoresis with three loci (Amy1, Amy2, and Gpi2). Genetic identities and the Fitch-Margoliash tree suggest differentiation into four regional groups: 1) a northwestern diploid group (northern France and northern Germany), 2) a northeastern diploid group (southern Germany, Upper Austria, northern Lower Austria, Poland, and Romania), 3) a central diploid group in southern Lower Austria corresponding to subspecies austriaca, and 4) a southern tetraploid group in Alpine areas of France, Germany, Switzerland, Austria, Italy, and Slovenia corresponding to subspecies laevigata. Geographically isolated diploid relic populations that are genetically depauperate are found in the NW and NE diploid groups. On the other hand, the diploid relic subspecies austriaca from the NE Prealps and Alps is highly variable. Subspecies laevigata appears to be a genetical autotetraploid with multiple origins involving several diploid progenitors (the NW diploids, subspecies austriaca and B. prealpina). Received April 6, 2001; accepted March 6, 2002 Published online: October 14, 2002 Addresses of the authors: Karin Tremetsberger (e-mail: k.tremetsberger@gmx.net), Christiane K?nig, Rosabelle Samuel, Tod F. Stuessy, Department of Higher Plant Systematics and Evolution, Institute of Botany, University of Vienna, Rennweg 14, A-1030 Vienna, Austria. Wilhelm Pinsker, Institute of Medical Biology, University of Vienna, Waehringerstra?e 10, A-1090 Vienna, Austria.     

Glacial in situ survival in the Western Alps and polytopic autopolyploidy in Biscutella laevigata L. (Brassicaceae)

Past climatic changes and especially the ice ages have had a great impact on both the distribution and the genetic composition of plant populations, but whether they promoted speciation is still controversial. The autopolyploid complex Biscutella laevigata is a classical example of polyploidy linked to glaciations and is an interesting model to explore migration and speciation driven by climate changes in a complex alpine landscape. Diploid taxa survived the last glacial maximum in several never-glaciated areas and autotetraploids are clearly dominant in the central parts of the Alps; however, previous range-wide studies failed to identify their diploid ancestor(s). This study highlights the phylogeographical relationships of maternal lineages in the Western Alps and investigates the polyploidy process using plastid DNA sequences (trnS-trnG and trnK-intron) combined with plastid DNA length polymorphism markers, which were transferable among Brassicaceae species. Twenty-one distinct plastid DNA haplotypes were distinguished in 67 populations densely sampled in the Western Alps and main lineages were identified by a median-joining network. The external Alps harboured high levels of genetic diversity, while the Central Alps contained only a subset of haplotypes due to postglacial recolonization. Several haplotypes were restricted to local peripheral refugia and evidence of in situ survival in central nunataks was detected by the presence of highly differentiated haplotypes swamped by frequent ones. As hierarchical genetic structure pointed to an independent evolution of the species in different biogeographical districts, and since tetraploids displayed haplotypes belonging to different lineages restricted to either the northern or the southern parts of the Alpine chain, polytopic autopolyploidy was also apparent in the Western Alps.     

PERMANENT GENETIC RESOURCES: Isolation of microsatellite loci from the endangered plant Sibara filifolia (Brassicaceae)

Sibara filifolia (Brassicaceae) is a federally endangered annual herb found on two of the California Channel Islands. Previous studies based on allozymes revealed little genetic variability on San Clemente Island. Nine polymorphic microsatellite loci were isolated from individuals on San Clemente Island. We found low levels of allelic variation (mean N_A = 2.3), with seven loci exhibiting significant deviations from Hardy–Weinberg equilibrium (P < 0.01) and 10 pairs of loci exhibiting significant linkage disequilibrium (P < 0.01). Most of the observed variability (mean H_O = 0.003) occurred among populations or in rare homozygous individuals.     

Fine-scale genetic structure and marginal processes in an expanding population of Biscutella laevigata L. (Brassicaceae)

Evolutionary processes acting at the expanding margins of a species' range are still poorly understood. Genetic drift is considered prevalent in marginal populations, and the maintenance of genetic diversity during recolonization might seem puzzling. To investigate such processes, a fine-scale investigation of 219 individuals was performed within a population of Biscutella laevigata (Brassicaceae), located at the leading edge of its range. The survey used amplified fragment length polymorphisms (AFLPs). As commonly reported across the whole species distribution range, individual density and genetic diversity decreased along the local axis of recolonization of this expanding population, highlighting the enduring effect of the historical colonization on present-day diversity. The self-incompatibility system of the plant may have prevented local inbreeding in newly found patches and sustained genetic diversity by ensuring gene flow from established populations. Within the more continuously populated region, spatial analysis of genetic structure revealed restricted gene flow among individuals. The distribution of genotypes formed a mosaic of relatively homogenous patches within the continuous population. This pattern could be explained by a history of expansion by long-distance dispersal followed by fine-scale diffusion (that is, a stratified dispersal combination). The secondary contact among expanding patches apparently led to admixture among differentiated genotypes where they met (that is, a reshuffling effect). This type of dynamics could explain the maintenance of genetic diversity during recolonization.     

珍稀濒危植物长蕊木兰种群的年龄结构与空间分布

珍稀濒危植物长蕊木兰为国家Ⅰ级保护植物。然而,由于受研究尺度和分析方法的限制,对其种群生态特征等方面仍不清楚。以云南高黎贡山原生的中山湿性常绿阔叶林4 hm2样地调查数据为基础,应用Ripley的L函数分析了长蕊木兰种群的年龄结构与空间分布格局。研究发现:(1)长蕊木兰种群的年龄结构为反"J"型,属稳定型种群。(2)长蕊木兰种群个体的空间分布格局与空间尺度关系密切,空间尺度小于75 m时为聚集分布,大于75 m时为随机分布。生境异质性在长蕊木兰种群空间分布格局的形成中可能发挥了重要的作用。(3)不同发育阶段个体的空间分布格局存在明显的差异,中树和小树阶段的分布格局在中、小尺度上呈聚集分布,在较大尺度上呈随机分布;大树阶段在整个空间尺度上均呈现随机分布。(4)长蕊木兰不同发育阶段的空间关联性主要表现为中、小尺度上的负相关,在较大尺度上则趋向于无关联。     

Genome-wide association to fine-scale ecological heterogeneity within a continuous population of Biscutella laevigata (Brassicaceae)

Gene flow, drift and selection can be detected through different signatures across the genome and the landscape. Genetic discontinuities along with their correlation to environmental features can be used to tease out isolation-by-distance and isolation-by-time from processes related to selection. Using spatial statistics (spatial autocorrelation methods, canonical correspondence analysis and partial Mantel tests) dealing with genome-wide amplified fragment length polymorphism (AFLP) under unlikely Hardy-Weinberg assumptions, this study investigates 124 individuals within a continuous population of the autopolyploid Biscutella laevigata (Brassicaceae). Fine-scale spatial genetic structure was strong and the mosaic-like distribution of AFLP genotypes was consistently associated with habitat factors, even when controlled for geographical distances. The use of multivariate analyses enabled separation of the factors responsible for the repartition of the genetic variance and revealed a composite effect of isolation by distance, phenological divergence and local adaptation to habitats characterised by different solar radiation regimes. These results suggest that the immigrant inviability barrier facilitated the maintenance of adapted subpopulations to distinct environmental conditions at the local scale.     

阿拉善左旗植物物种多样性空间分布特征

通过对内蒙古自治区阿拉善左旗的植被样方调查,研究干旱荒漠地区植物群落物种多样性的梯度变化和空间分布特征。通过对样方数据的群落生活型构成、物种丰富度、α多样性、β多样性分析,结合CCA排序和地统计方法,结果表明：（1）在该区域植物物种的α多样性和β多样性均起伏较大。（2）草本植物的丰富度由西向东呈增高趋势,而灌木的丰富度则呈降低趋势;随纬度的增加,草本植物丰富度呈现下降趋势,而灌木丰富度则显现出上升趋势。但是,38&#176;～39．2&#176;N之间出现了一个灌木和草本物种丰富度都相对比较低的异常区域。（3）α多样性与经度正相关,但与纬度存在负相关关系。β多样性显示,随着经度的增加,自西向东样方问物种构成的相似性降低,物种替代速率升高。随着纬度的增加,群落组成呈现逐渐单一化的趋势。（4）Shannon—Wiener指数和Shimpson指数的CO／（CO＋Cl）都在0．25—0．75之间,α多样性为中等空间相关性。CCA分析的结果表明,地理因素对于物种多样性有显著的影响,而且经度的影响大于海拔的影响。研究阿拉善左旗荒漠区植物物种多样性的梯度变化和空间分布特征,为认识和保护荒漠地区生物多样性资源提供了理论依据和实践基础。     

Conservation of genetic diversity in the endangered plant Eriogonum ovalifolium var. vineum (Polygonaceae)

The purpose of his research was to describethe organization of genetic variation in thefederally endangered plant taxon Eriogonumovalifolium var. vineum using allozymes. Such information can help prioritize sites andmanagement choices for capturing andmaintaining genetic variation and can reducethe number of populations necessary to committo conservation, thus reducing costs andconflicts with competing land uses. Information on genetic diversity patterns alsoprovides insight into evolutionary anddemographic history of a taxon which canprovide means to assess future risk of erosionof diversity. Similar to other Eriogonumovalifolium varieties, E. ovalifoliumvar. vineum is diverse atboth the taxon (proportion of polymorphic loci[P] = 0.55 and alleles per locus [A] =5.45 [SD = 2.5]) and population (P = 0.56 [SD =0.11] and A = 2.68 [SD = 0.35]) levels. Gene diversity (H _e) withinpopulations averaged 0.19 (SD = 0.03). Wefound some evidence for limited clonalreproduction within populations. Populationswere moderately differentiated from one another(_S = 0.14) and showed moderatedeviations from Hardy-Weinberg equilibriumwithin populations (f = 0.14 and F =0.19). Mean Nei's genetic distance (D)among all pairs of populations was 0.02. Populations with high levels of multiplemeasures of genetic diversity, high levels ofdifferentiation, and low levels of apparentinbreeding are suggested as conservationpriorities.     

Genetic diversity and similarity in the Barbarea vulgaris complex (Brassicaceae)

Two subspecies of Barbarea vulgaris are taxonomically recognized as ssp. vulgaris and ssp. arcuata. In addition, two types of Barbarea vulgaris ssp. arcuata occurs in Denmark. The G‐type is resistant to an herbivorous flea beetle (Phyllotreta nemorum) whereas the P‐type is susceptible. A previous study suggested that the P‐type evolved by a loss of resistance from a resistant progenitor. We analyzed the genetic relatedness among eight Barbarea taxa: B. vulgaris spp. vulgaris, B. vulgaris ssp. arcuata G‐ and P‐types, hybrids between the types, B. verna, B. intermedia, B. stricta, B. orthoceras and B. australis, using AFLP and SSR markers. A clear distinction between the G‐ and P‐types was revealed. Both were distinct from B. vulgaris ssp. vulgaris, the G‐type less so than the P‐type. Barbarea verna and B. intermedia formed unambiguous clusters, whereas the remaining taxa produced less discrete groupings. Possible evolutionary scenarios for flea‐beetle resistance and susceptibility are discussed, including lineage sorting from a polymorphic ancestral population, and de novo loss of resistance in the P‐type of B. vulgaris ssp. arcuata.     

濒危植物元宝山冷杉的遗传多样性研究

元宝山冷杉(Abies yuanbaoshanensis)是仅分布于广西融水县元宝山的珍稀濒危物种。本研究采用AFLP分子标记分析了元宝山冷杉种群的遗传多样性。取元宝山冷杉种群43棵植株作为研究材料,4对引物组合用于AFLP扩增,共得到261条DNA扩增带。分析结果：元宝山冷杉种群的多态位点百分比率为50．96％,Nei's基因多样性为0．1510,Shannon多样性指数为0．1735。用NTSYS软件计算各个样品间的相似性系数,并用UPGMA法基于相似性系数进行了聚类分析。研究结果表明：元宝山冷杉种群的遗传多样性水平低,种群内个体间的相似性系数很大,无明显亚种群间的遗传分化。要保护或保存这个物种的遗传资源,建议应该选择种群内尽可能多的个体进行保护。     

Three new Spanish species of Biscutella L. (Brassicaceae) and remarks on B. valentina (L.) Heywood

Three new species are described in Biscutella sect. Biscutella from the central-eastern mountains of the Iberian Peninsula: B. bilbilitana Mateo & M.B. Crespo, B. segurae Mateo & M.B. Crespo, and B. conquensis Mateo & M.B. Crespo. They are characterized morphologically, ecologically and chorologically. Data separating them from B. valentina (L.) Heywood, a taxon to which they sometimes have been referred, are also reported.     

珍稀濒危植物天目木兰（Magnolia amoena）遗传多样性的RAPD分析

运用随机扩增多态DNA(RAPD)技术,对天目木兰(Magnolia amoena)居群的遗传多样性进行了研究．从40个10-mer随机引物中筛选出14个能得到清晰、稳定扩增带的引物进行扩增,14个引物共检测了94个位点,其中多态性位点为23,占24．4％,计算了12个居群之间的遗传相似度和遗传距离,并运用UPGMA法进行了聚类分析,结果显示相同严地个体间(居群内)的遗传距离较小,遗传多样性水平很低;不同产地个体间(居群间)遗传距离较大,遗传多样性水平较前者高,即天目木兰个体间遗传多样性水平与它的地理分布有关,天目木兰总体较低的遗传多样性是导致它濒危的原因之一。     

黄山花楸种群遗传多样性研究

利用RAPD技术,对黄山花楸(Sorbus amabilis)自然分布区的13个自然种群的遗传多样性进行了研究,从40个10碱基随机引物中筛选出能产生稳定多态性标记的引物14个,共扩增出105个位点,其中多态性位点30个,占28．6％,应用UPGMA法和Neighbor-Joining法对遗传距离进行聚类分析构建树系图。结果表明,黄山花楸自然种群具有较低的遗传多样性,对环境变化的适应能力较差;其种群间的遗传差异与其地理分布有关;黄山花楸自身的特殊进化历史和人为砍伐以及自然灾害(火灾、病虫害等)和小种群的遗传漂变作用是黄山花楸遗传多样性水平低的主要原因,也是其濒危的主要原因。     

The small-scale spatial distribution of an invading moth

We studied the spread of a small leaf-mining moth [Phyllonorycter leucographella (Zeller), Gracillariidae] after its accidental introduction into the British Isles. At large geographical scales, previous work had shown the spread to be well described by a travelling weve of constant velocity. Here, we report the pattern of spread at scales of 1 km². By locating all bushes of the insect's foodplant (Pyracantha spp.) within 1-km² quadrats, the precise pattern of colonisation at finer spatial scales could be established. Where the 1-km² site was colonised by moths from the main advancing front, no spatial pattern in the order that bushes were infested was found. If the source of colonisation was a single or small group of infested plants within the site, there was some evidence that nearby plants were colonised first. We found no evidence of population turnover after colonisation. We interpret the results in terms of a two-stage model of invasion that produces different patterns at small and large geographical scales.     

The small-scale spatial distribution of male and female plants

Summary There are many reports of dioecious flowering plants with positive spatial association of gender, such that males and females tend to occur at different sites. This pattern has been interpreted as demonstrating environmenal heterogeneity, the two sexes having different habitat preferences. However, it is not clear: (a) to what extent the literature reflects the greater likelihood of publishing striking positive results; and (b) whether the association of stems of like gender within habitats is caused by differences in microhabitat preference rather than by vegetative ramification. We surveyed natural populations of ten sexually heteromorphic species in southern Quebec, and found that five showed positive association and four showed no association between the genders. With one exception (Silene cucubalus) these results appeared to reflect the presence or absence of vegetative ramification. We conclude that the demonstration of spatial association cannot be used to infer the existence of microenvironmental heterogeneity except in species with no capacity for vegetative ramification. The tenth species, Silene alba, showed negative association, with an excess of neighbouring pairs of unlike gender, but we were unable to reproduce this phenomenon experimentally.     

Homogeneity at nuclear microsatellite loci masks mitochondrial haplotype diversity in the endangered fanshell pearlymussel (Cyprogenia stegaria)

We report on multiple patterns of differentiation and connectivity in the fanshell pearlymussel (Cyprogenia stegaria), based on different markers. Knowledge of genetic variation and genetic connectivity among remaining populations of this federally endangered species is needed to initiate implementation of the species recovery plan. We collected tissue samples from 96 specimens from the Green, Rolling Fork, and Licking Rivers, tributaries to the Ohio River, and the Clinch River, a tributary to the Tennessee River, providing broad coverage of the current distributional range of the species. Results from 7 nuclear DNA microsatellite markers suggested minimal population-level differentiation, whereas a mitochondrial DNA (mtDNA) marker (ND1) exhibited significant differentiation between C. stegaria in the Clinch River and the Ohio River populations. The ND1 data also confirm the existence of 2 distinct mtDNA lineages in the genus that transcends species boundaries. Further analyses suggest that the disproportionally strong signal from 2 very divergent ND1 lineages possibly masks finer-grained structure in the Ohio River population, based on one of the mtDNA lineages only. We recommend further sampling to confirm the absence of one lineage from the upper Clinch River drainage and suggest that provisional management guidelines should limit reciprocal exchanges among C. stegaria populations from the Clinch River and those in the Ohio River system.     

濒危植物资源冷杉遗传多样性研究

资源冷杉(Abies ziyuanensis)属松科(Pinaceae)冷杉属(Abies),是国家一级保护的濒危植物。利用RAPD分子标记对分布于广西资源县银竹老山和湖南省炎陵县大院的2个资源冷杉种群共54株进行了遗传多样性分析。用20个随机引物共扩增到126个位点,其中66个位点是多态性的,总的多态位点百分率为52.4％,在两个种群分别为33.6％和31。2％。Nei's基因多样性为0.312,Shannon信息指数为0.475,两个种群的遗传分化系数Gst为0.455。基于Nei's遗传距离进行了UPGMA聚类分析,结果两种群的样品彼此明显区分开来。分析结果表明资源冷杉种群内的遗传多样性较低,但两种群间的遗传分化明显。需要同时重视两分布地的资源冷杉的保护工作。