Genetic diversity and dispersal of the moss Sarconeurum glaciale on Ross Island, East Antarctica

The extent of genetic variation and dispersal mechanisms were investigated over short distances of 1–100 m, and up to 3 km, by the random amplified polymorphic DNA (RAPD) technique, for the moss Sarconeurum glaciale , at three locations on Ross Island, Antarctica. At Arrival Heights, genetic variation occurred within single colonies, and the relationships between clumps indicated that they were dispersed down small, meltwater drainage channels by water. The genetic similarities between the colonies from Arrival Heights and others from Scott Base and Crater Hill, a few km away, together with the prevailing wind direction and absence of this moss in the intervening snow-covered area, suggested longer-distance dispersal by wind. Overall, the Ross Island samples appeared to form a single, polymorphic population that was distinct from another population at Canada Glacier, 110 km distant. Somatic mutation, rather than immigration of genetically different propagules from elsewhere, appeared to be the most probable cause of genetic variability in these haploid, vegetatively reproducing Antarctic moss populations. Initiation of recolonization of S. glaciale across a dirt track at Arrival Heights was also investigated by RAPDs, to investigate how regrowth of mosses in disturbed areas occurred in the extreme environment of Antarctica.     

A survey of metal concentrations in higher plants, mosses, and lichens collected on King George Island in 1988

Antarctica is considered to be one of the least polluted regions on earth, and therefore, it is important to survey and control the level of contamination. Antarctic vegetation is very sparse and is essentially restricted to seashore oases and nunataks. Therefore, any data concerning metal levels in plants and lichens are of crucial value for this area. Our first goal was to determine metal concentrations in two higher plants and the most dominant species of mosses and lichens collected in 1988. We then compared the results of our survey with recent studies employing similar methodology. In our study, Cr, Cu, Fe, Ni, Pb, V, and Zn concentrations in mosses, C. quitensis and D. antarctica were also higher than typical values for mosses and vascular plants from unpolluted areas indicating anthropogenic influence. Mosses were determined to be better bioindicators of metals than lichens. Hg concentrations in mosses were significantly higher than those in shoots of C. quitensis and D. antarctica. Increases in Cr, Pb, and V concentrations over time were observed in moss when concentrations from samples collected in 1988 were compared with more recent data from other studies. Our results for King George Island may apply at least to all the maritime Antarctic where climate and plant communities are similar.     

Isolation of Bacillus thuringiensis from Northern Victoria Land, Antarctica

Sixty-two samples of Antarctic soils, mosses, penguin guano, algae and lichens were examined for the presence of aerobic endospore-forming bacteria; 36 samples (58%) yielded such organisms, and two samples from Edmonson Point (74 degrees 21'S 165 degrees 08'E) and one sample from Apostrophe Island (73 degrees 32'S 167 degrees 24'E), northern Victoria Land, yielded strains of Bacillus thuringiensis. Further isolations from two of the samples, appreciable variation in biotypes among the strains, failure of the strains to grow on routine B. thuringiensis media, and the fact that one of the sampling sites is very rarely visited by humans, suggest that the organisms were living in these soils rather than being chance contaminants. A representative strain, from Apostrophe Island, was identified as serovar pirenaica (H57).     

Spatial and temporal variability across life's hierarchies in the terrestrial Antarctic

Antarctica and its surrounding islands lie at one extreme of global variation in diversity. Typically, these regions are characterized as being species poor and having simple food webs. Here, we show that terrestrial systems in the region are nonetheless characterized by substantial spatial and temporal variations at virtually all of the levels of the genealogical and ecological hierarchies which have been thoroughly investigated. Spatial variation at the individual and population levels has been documented in a variety of genetic studies, and in mosses it appears that UV-B radiation might be responsible for within-clump mutagenesis. At the species level, modern molecular methods have revealed considerable endemism of the Antarctic biota, questioning ideas that small organisms are likely to be ubiquitous and the taxa to which they belong species poor. At the biogeographic level, much of the relatively small ice-free area of Antarctica remains unsurveyed making analyses difficult. Nonetheless, it is clear that a major biogeographic discontinuity separates the Antarctic Peninsula and continental Antarctica, here named the 'Gressitt Line'. Across the Southern Ocean islands, patterns are clearer, and energy availability is an important correlate of indigenous and exotic species richness, while human visitor numbers explain much of the variation in the latter too. Temporal variation at the individual level has much to do with phenotypic plasticity, and considerable life-history and physiological plasticity seems to be a characteristic of Antarctic terrestrial species. Environmental unpredictability is an important driver of this trait and has significantly influenced life histories across the region and probably throughout much of the temperate Southern Hemisphere. Rapid climate change-related alterations in the range and abundance of several Antarctic and sub-Antarctic populations have taken place over the past several decades. In many sub-Antarctic locations, these have been exacerbated by direct and indirect effects of invasive alien species. Interactions between climate change and invasion seem set to become one of the most significant conservation problems in the Antarctic. We conclude that despite the substantial body of work on the terrestrial biodiversity of the Antarctic, investigations of interactions between hierarchical levels remain scarce. Moreover, little of the available information is being integrated into terrestrial conservation planning, which lags far behind in this region by comparison with most others.     

The genetic structure of endangered populations in the Cranberry Fritillary,Boloria aquilonaris (Lepidoptera,Nymphalidae): RAPDs vs allozymes

The genetic population structure of the Cranberry Fritillary Boloria aquilonaris was studied using both RAPDs (random amplified polymorphic DNA) and allozymes. In Belgium, B. aquilonaris has a naturally fragmented distribution that has been accentuated due to human activity during the last century. The genetic population structure of this butterfly was analysed at the regional (several Ardenne uplands) and at the landscape level (several populations within an Ardenne upland). Both population genetic markers confirmed results from a previous CMR study at the landscape scale. At the regional scale however, important incongruences were observed between RAPDs and allozymes. The average gene diversity for the RAPD data was twice that of the allozyme data. The degree of population subdivision was also much greater for RAPDs than for allozymes. The UPGMA clusters produced by each of these markers differed significantly. We believe that, given the higher rate of mutation of RAPDs and the greater number of loci assayed by this method, RAPDs reveal a more accurate and recent population genetic structure than allozymes.     

Genetic relationships among Native American maize accessions of the Great Plains assessed by RAPDs

Genetic variation among 15 accessions of Native American maize from the Great Plains was investigated using random amplified polymorphic DNA (RAPD). RAPDs revealed very high levels of polymorphism among accessions. Banding patterns ranged in percentage polymorphism from 46.7% to 86.2% with an overall mean of 70.7% for the primers analyzed. The construction of genetic relationships using cluster analysis and principal coordinates analysis revealed that RAPDs are successful in confirming hypothesized relationships and in identifying misclassified specimens. Furthermore, the phenogram fails to reveal a strong correspondence between genetic relationships and the geographical position of Native Americans prior to contact. This provides support for the hypothesis that multiple introductions of maize into the Great Plains via trade may have resulted in the great morphological variation found among accessions in the region. Based on these data, it is unlikely that a separate Great Plains race of maize can be distinguished. In general, we conclude that RAPDs are potentially very useful in organizing seed collections and understanding intraspecific genetic differentiation. Received: 10 March 1999 / Accepted: 25 March 1999     

采用RAPD和ISSR标记研究古尔班通古特沙漠东南缘梭梭种群的遗传结构

梭梭(Haloxylon ammodendron(CA Mey.)Bunge)是一种沙漠旱生优势树种,具有重要的生态和经济价值,然而,我们对梭梭种群的遗传多样性和遗传结构所知甚少.本文采用RAPD和ISSR标记对来自古尔班通古特沙漠东南缘的4个天然梭梭种群的遗传多样性和遗传结构进行了检测.5个RAPD引物和8个ISSR引物分别扩增出61和195条带,多态性位点比率分别为83.6%和89.7%,Shannon信息指数分别为0.333和0.367,RAPD和ISSR分析均表明梭梭种群的遗传多样性水平较高.利用分子方差分析(AMOVA)研究梭梭种群的遗传结构,结果表明,大部分遗传变异存在于种群内,通过RAPD分析发现138.2%的遗传变异发生在种群内;通过ISSR分析发现89.4%的遗传变异发生在种群内;而种群间的遗传分化很小.通过RAPD标记没有检测到种群间的遗传分化,ISSR分析表明10.6%的遗传变异发生在种群内.我们推测梭梭种群较高的遗传多样性水平可能源于对异质、高胁迫环境的长期适应,但还需要进一步的研究加以证实.种群间遗传分异低的主要原因是种群间存在强大的基因流.     

Radiocarbon bomb spike reveals biological effects of Antarctic climate change

The Antarctic has experienced major changes in temperature, wind speed and stratospheric ozone levels during the last 50 years. However, until recently continental Antarctica appeared to be little impacted by climate warming, thus biological changes were predicted to be relatively slow. Detecting the biological effects of Antarctic climate change has been hindered by the paucity of long‐term data sets, particularly for organisms that have been exposed to these changes throughout their lives. We show that radiocarbon signals are preserved along shoots of the dominant Antarctic moss flora and use these to determine accurate growth rates over a period of several decades, allowing us to explore the influence of environmental variables on growth and providing a dramatic demonstration of the effects of climate change. We have generated detailed 50‐year growth records for Ceratodon purpureus and three other Antarctic moss species using the 1960s radiocarbon bomb spike. Our growth rate and stable carbon isotope (δ¹³C) data show that C. purpureus’ growth rates are correlated with key climatic variables, and furthermore that the observed effects of climate variation on growth are mediated through changes in water availability. Our results indicate the timing and balance between warming, high‐wind speeds and elevated UV fluxes may determine the fate of these mosses and the associated communities that form oases of Antarctic biodiversity.     

Effects of reforestation methods on genetic diversity of lodgepole pine: an assessment using microsatellite and randomly amplified polymorphic DNA markers

 We examined the effects of different methods of forest regeneration on the genetic diversity of lodgepole pine (Pinus contorta var ‘latifolia’) using two different DNA-based molecular markers [randomly amplified polymorphic DNA (RAPDs) and microsatellites or simple sequence repeats (SSRs)]. Genetic diversity was estimated for 30 individuals in each of four populations for the following three stand types: (1) mature lodgepole pine (>100 years); (2) 20- to 30-year-old harvested stands left for natural regeneration; (3) 20- to 30-year-old planted stands (4 stands of each type); and one group of 30 operationally produced seedlings. There was no significant effect of stand type on expected heterozygosity, although allelic richness and diversity were much higher for SSRs than for RAPDs. Expected heterozygosity ranged from 0.39 to 0.47 based on RAPDs and from 0.67 to 0.77 based on SSRs. The number of alleles per locus for SSRs ranged from 3 to 34 (mean 21.0), and there was a significant relationship between sequence repeat length and the number of alleles at a locus. Both marker types showed that over 94% of the variation was contained within the populations and that the naturally regenerated stands sampled had lower (not significant) expected heterozygosity than the planted or unharvested stands. The group of seedlings (assessed by RAPDs only) had expected heterozygosity and allele frequencies similar to those of the unharvested stands. Genetic distance measures were higher than obtained previously in the species using isozyme markers. There was no correlation between the two marker types for pair-wise genetic distances based on populations analyzed by both methods. Pair-wise genetic distance measures and an ordination of allele frequencies for both marker types showed little effect of geographic location or stand type on genetic similarity. Received: 18 August 1998 / Accepted: 17 September 1998     

Evaluation of the extent of genetic variation in mahoganies (Meliaceae) using RAPD markers

Despite the economic importance of mahoganies (Meliaceae) little is known of the pattern of genetic variation within this family of tropical trees. We describe the application of a polymerase chain reaction (PCR)-based polymorphic DNA assay procedure random amplified polymorphic DNAs (RAPDs) to assess the extent of genetic variation between eight mahogany species from four genera. Pronounced genetic differentiation was found between the species and genera. There was a clear separation of Cedrela odorata from the other species, with 95% of the variable amplification products differing, whereas Lovoa trichilioides, Khaya spp. and Swietenia spp. were more closely grouped. These results are consistent with the current taxonomic viewpoint. A number of markers were found to be diagnostic for particular species, which could be of value in determining the status of putative hybrids. The application of RAPDs to the study of genetic variation in mahoganies is discussed in the context of developing genetic conservation and improvement strategies for these species.     

Extent and patterns of RAPD variation in landraces and cultivars of rye (Secale cereale L.) from northern Europe

Little is known about the extent and patterns of distribution of RAPD diversity in outcrossing species. This study is the first step in using RAPD markers to quantify the amount and distribution of genetic variation within and between accessions of 9 landraces and 3 cultivars of cultivated rye from Northern Europe. A high level of RAPD variation was detected, demonstrating the utility of RAPDs for genetic characterisation in rye. The results show that: (1) landraces and improved cultivars maintain roughly the same high levels of RAPD variation, (2) landraces from Norway, Germany and Finland showed the lowest level of variation, probably because of a small amount of seeds from the original samples, (3) most of the RAPD variation was found within rather than between the accessions, which is consistent with the pattern expected for a cross pollinated crop. Both the cluster and the principal coordinates analyses displayed the same pattern of genetic relationship among the accessions studied.     

Tracking down human contamination in ancient human teeth

DNA contamination arising from the manipulation of ancient calcified tissue samples is a poorly understood, yet fundamental, problem that affects the reliability of ancient DNA (aDNA) studies. We have typed the mitochondrial DNA hypervariable region I of the only 6 people involved in the excavation, washing, and subsequent anthropological and genetic study of 23 Neolithic remains excavated from Granollers (Barcelona, Spain) and searched for their presence among the 572 clones generated during the aDNA analyses of teeth from these samples. Of the cloned sequences, 17.13% could be unambiguously identified as contaminants, with those derived from the people involved in the retrieval and washing of the remains present in higher frequencies than those of the anthropologist and genetic researchers. This finding confirms, for the first time, previous hypotheses that teeth samples are most susceptible to contamination at their initial excavation. More worrying, the cloned contaminant sequences exhibit substitutions that can be attributed to DNA damage after the contamination event, and we demonstrate that the level of such damage increases with time: contaminants that are >10 years old have approximately 5 times more damage than those that are recent. Furthermore, we demonstrate that in this data set, the damage rate of the old contaminant sequences is indistinguishable from that of the endogenous DNA sequences. As such, the commonly used argument that miscoding lesions observed among cloned aDNA sequences can be used to support data authenticity is misleading in scenarios where the presence of old contaminant sequences is possible. We argue therefore that the typing of those involved in the manipulation of the ancient human specimens is critical in order to ensure that generated results are accurate.     

Genetic variation at allozyme and RAPD loci in sessile oak Quercus petraea (Matt.) Liebl.: the role of history and geography

The nuclear genetic variation within and among 21 populations of sessile oak was estimated at 31 RAPD loci in conjunction with previous estimates of variation at eight allozyme loci. The aim of the study was to assess the relative role of isolation-by-distance and postglacial history on patterns of nuclear variation. Because of its small effective population size and maternal transmission, the chloroplast genome is a good marker of population history. Both kinds of nuclear variation (RAPD and allozyme) were therefore compared, first, to the geographical distances among populations and, secondly, to chloroplast DNA restriction polymorphism in the same populations. Multiple Mantel tests were used for this purpose. Although RAPDs revealed less genetic diversity than allozymes, levels of genetic differentiation ( G _ST) were identical. The standard genetic distance calculated at all RAPD loci was correlated with geographical distances but not with the genetic distance calculated from chloroplast DNA data. Conversely, allozyme variation was correlated with chloroplast DNA variation, but not with geography. Possibly, divergent selection at two allozyme loci during the glacial period could explain this pattern. Because of its greater number of loci assayed, RAPDs probably provided a less biased picture of the relative role of geography and history.     

Microbiological Evaluation of Suture Items Before Radiation Sterilization

Microbiological contamination levels of suture samples taken at various stages of the manufacturing process in a new hygienically controlled plant were determined by employing a membrane filturation technique. Both raw material and materials handled manually in the production process were tested to assess the effect of manual handling on the product contamination level. Evaluation of the efficacy of contamination control, however, was directed primarily to the finished, packaged products, just prior to the processing with cobalt 60. The suture material for testing was divided into two groups, namely, wet and dry products, the wet being packaged in a special "tubing" fluid consisting mainly of isopropyl alcohol. Initial contamination results are reported as the average of values obtained on the test day and the preceding 9 consecutive production days. A total of 1,787 suture samples tested in the dry group showed daily averages varying between 2.1 and 14.8 contaminants per suture. The 2,980 wet-packaged suture samples tested gave daily averages varying from 0.7 to 4.2 contaminants per suture. The highest values obtained for an individual suture were 400 for the dry and 89 for the wet. Identification studies of the contaminants revealed that fungi predominated. Most of the bacterial contaminants proved to be spore-forming rods.     

Genetic variation in Antarctic populations of the moss Sarconeurum glaciale

Sixty-six isolates of the moss Sarconeurum glaciale were collected from sites in continental Antarctica at Ross Island, southern Victoria Land and the Vestfold Hills. Genetic variation within and among the populations was estimated using isozymes and random amplified polymorphic DNA (RAPD) technology. Isozyme results only reproducibly showed variation between the populations with one enzyme; RAPDs indicated significantly higher levels of genetic variability within and among the Vestfold Hills samples than in the Ross Sea region samples. A dendrogram produced from the RAPD bands suggested that the Ross Island and southern Victoria Land samples form one population, and those from the Vestfold Hills form a separate and more variable population, possibly resulting from separate colonisation events on the continent. Received: 15 March 1996 / Accepted: 1 May 1997     

羊草种群遗传分化的RAPD分析Ⅱ.RAPD数据的统计分析

对松嫩草原上分布的灰绿型和黄绿型羊草9个种群进行了15个引物的RAPD分析,统计结果表明,两类种群的扩增片段数和多态位点比率明显不同,黄绿型种群低于灰绿型,其值分别＜90与＞100,＜50％与＞70％,比较了7种不同统计方法据RAPD表型或基因型频率估算的种群遗传多样性,几种统计结果都揭示,黄绿型种群低于灰绿型种群,用F1s值矫正种群对Hardy-Weinberg平衡的偏离后,估算等位基因频率,通过Shannon指数和Nei指数估计羊草种群间分化分别为37．6％和35．7％,高于等位酶的分析,讨论比较了等位酶和RAPD分析结果的异同。     

Terrestrial contamination in Apollo lunar samples

The Apollo lunar samples were seen to offer a unique opportunity in the search for extraterrestrial organic matter without the ambiguity surrounding meteorite analysis due to their unknown contamination histories. The recognition that only a small amount of indigenous organic material was likely to be present in lunar samples combined with the extreme sensitivity of organic analysis methods made it clear that this opportunity could be realized only by carefully controlling the collection, processing, and analysis of the samples in order that they might remain free of significant levels of contamination. The contamination control procedures adopted are described and the analytical evidence obtained throughout the program on potential contamination sources is presented. The organic contaminants actually found in the lunar samples by the various investigators are summarized. It is shown that the program succeeded in providing investigators with samples containing less than 0.1 ppm total contamination.     

High levels of genetic variability in the moss Ceratodon purpureus from continental Antarctica, subantarctic Heard and Macquarie Islands, and Australasia

The random amplified polymorphic DNA (RAPD) technique, and DNA sequencing of the conserved nuclear ribosomal DNA internal transcribed spacer region (ITS1-5.8S-ITS2), have been used to assess levels of genetic diversity in the moss Ceratodon purpureus from several locations in Australasia, subantarctic Heard and Macquarie Islands, and continental Antarctica. Populations from Heard and Macquarie Islands and from Antarctica maintain high levels of genetic variation. Both within- and among-colony variation were observed at these locations. DNA sequence analysis showed that samples from the Ross Sea region of Antarctica were most closely related to colonies from Casey and Macquarie Island, and that one colony from Heard Island was most closely related to one from Europe. DNA sequence data separated two Australian populations from the Antarctic and subantarctic group on a dendrogram. Detailed RAPD analysis of a single colony from continental Antarctica demonstrated that mutation probably causes the high variability observed in this moss. DNA sequencing and RAPD analysis are complementary techniques for genetic investigation of Antarctic moss populations.Jenny Ninham was an integral part of this research team for several years. Unfortunately she did not live to see these results published.     

Ancient DNA analysis of human populations

The use of ancient DNA (aDNA) in the reconstruction of population origins and evolution is becoming increasingly common. The resultant increase in number of samples and polymorphic sites assayed and the number of studies published may give the impression that all technological hurdles associated with aDNA technology have been overcome. However, analysis of aDNA is still plagued by two issues that emerged at the advent of aDNA technology, namely the inability to amplify a significant number of samples and the contamination of samples with modern DNA. Herein, we analyze five well-preserved skeletal specimens from the western United States dating from 800-1600 A.D. These specimens yielded DNA samples with levels of contamination ranging from 0-100%, as determined by the presence or absence of New World-specific mitochondrial markers. All samples were analyzed by a variety of protocols intended to assay genetic variability and detect contamination, including amplification of variously sized DNA targets, direct DNA sequence analysis of amplification products and sequence analysis of cloned amplification products, analysis of restriction fragment length polymorphisms, quantitation of target DNA, amino acid racemization, and amino acid quantitation. Only the determination of DNA sequence from a cloned amplification product clearly revealed the presence of both ancient DNA and contaminating DNA in the same extract. Our results demonstrate that the analysis of aDNA is still an excruciatingly slow and meticulous process. All experiments, including stringent quality and contamination controls, must be performed in an environment as free as possible of potential sources of contaminating DNA, including modern DNA extracts. Careful selection of polymorphic markers capable of discriminating between ancient DNA and probable DNA contaminants is critical. Research strategies must be designed with a goal of identifying all DNA contaminants in order to differentiate convincingly between contamination and endogenous DNA.