Patterns of genotypic diversity suggest a long history of clonality and population isolation in the Australian arid zone shrub Acacia carneorum

For plants capable of both sexual and clonal reproduction, the relative frequency of these reproductive modes is influenced by genetic and ecological factors. Acacia carneorum is a threatened shrub from the Australian arid zone that occurs as a set of small, spatially isolated populations. Sexual reproduction appears to be very rare: despite regular flowering, only two populations set seed. It is not known whether this reflects an ancient pattern, or results from rapid land use changes following arrival of Europeans in the region 150 years ago. We assessed genotypic variation throughout the range of A. carneorum using AFLP markers, to elucidate the relative importance of clonal and sexual reproduction in this species’ history. Clonal diversity (CD) within populations ranged from 0 to 0.820 (mean CD = 0.270, SE = 0.094), but the relative abundances of genets were typically highly skewed. On average, the two fruiting populations had higher CD (mean CD = 0.590, SE = 0.265) than non-fruiting populations (mean CD = 0.179, SE = 0.077) (t = 2.315, p = 0.049), but most populations contained multiple genets. All genets were population-specific, and there was substantial divergence among populations (Φ _ST = 0.690), implying a long history of isolation. We conclude that clonality has predominated in A. carneorum populations, with occasional sexual recruitment, and that current failure of most populations to set seed likely reflects both a long history of asexual reproduction and effects of habitat disturbance. Conservation of this species may benefit from translocations to increase genotypic diversity within populations.     

Assessing genetic structure in a rare clonal eucalypt as a basis for augmentation and introduction translocations

Genetics can provide information that is critical for planning translocations for conservation, such as levels of diversity and divergence of target and source populations. For clonal plants, assessing population characteristics (size, diversity, mortality, gene flow) that influence conservation values also requires identification of different genetic individuals. We used 12 microsatellite markers to guide germplasm source recommendations for augmentation and introduction translocations to conserve the critically endangered Eucalyptus cuprea that occurs in fragmented populations in the semi-arid shrublands of Western Australia. Ramet clumps with identical multilocus genotypes were identified in all populations but clonal richness (R = 0 ? 0.86) and heterogeneity (D = 0 ? 0.98) varied among populations. Genetic diversity was low to moderate in all populations (mean H _o = 0.61, mean A = 3.78) and did not differ significantly between localities. There was evidence of inbreeding in some populations but outcrossing (t _m = 0.495) in the small number of families available for study (N = 4) and genotypic diversity of the larger extant populations suggest the generation of novel genotypes is a component of the reproductive strategy. Most diversity was within populations and differentiation among populations was moderate (F _ST = 0.100) suggesting mixing of source population for translocation is unlikely to lead to outbreeding depression. Principal Co-ordinate and Bayesian analyses indicated the Northern population is distinct from Central/Southern populations. We recommend use of mixed germplasm to conserve the moderate diversity characterising larger remnant populations and to enable the production of recombinants through sexual reproduction. But given seed availability and the distinction of the Northern population, an initial precautionary approach to a translocation proposed for south of the geographical range may be to source germplasm from the Central/Southern locality.     

Clonal diversity and spatial genetic structure of Potamogeton pectinatus in managed pond and river populations

Higher levels of genetic diversity of river macrophytes are expected in downstream parts because of potential accumulation of various genotypes from upstream sites. We assessed the clonal diversity and spatial genetic structure of fennel pondweed (Potamogeton pectinatus or Stuckenia pectinata) populations with emphasis on the estimation of dispersal via clonal propagules along a river in connection to upstream ponds. We analysed genetic diversity of 354 plant shoots sampled in 2005 and 2006 at three pond and five river sites in the Woluwe river catchment (Belgium). Nine microsatellite DNA markers revealed 88 genets of which 89% occurred in only one site. Clonal propagule dispersal was detected up to 10 km along the river. Few multilocus genotypes were repeatedly present along a major part of the river indicating vegetative spread. Populations of ponds contained a higher amount of clonal diversity, indicating the importance of local seed recruitment. A fine-scaled spatial genetic structure indicated that most seedling recruitment occurred at a distance <5 m in pond populations whereas clones in river sites were unrelated and showed no spatial autocorrelation. The clonal diversity decreased along the river from upstream to downstream due to establishment of few large clones.     

High level of genetic variation within clonal orchid Goodyera repens

Type of reproduction has an important effect on the maintenance of particular populations and species persistence in time and space. This trait significantly influences the ecological and genetic structure of populations, and in consequence the evolution of species. The primary objectives of this study were: to estimate genetic diversity within and among populations of clonal species Goodyera repens from different populations in northeastern Poland, and to amount factors shaping the genetic structure of this orchid. Based on 451 rosettes of G. repens from 11 localities in northeastern Poland, we conducted a genetic population analysis using allozymes. We included information on population size, flowering, fruit set and seed dispersal to elucidate their influences on genetic diversity of this species. Populations differed according to demographic properties. The majority of seeds (86.4–94.8 %) were found at a distance of 0.2 m. We observed a high level of genetic (P _PL = 50 %, A = 1.68, H _O = 0.210, H _E = 0.204) and genotypic diversity (G = 163, G/N _S = 0.66, G _U = 30.2 %), and low but statistically significant genetic differentiation among populations (F _ST = 0.060; P < 0.001). We suggest that the genetic diversity of G. repens is mainly an effect of the abundance of pine and spruce forest communities suitable for this species in NE Poland and the high level of sexual reproduction.     

Population genetic structure of wild daffodils (Narcissus pseudonarcissus L.) at different spatial scales

We studied the population genetic and clonal structure of the endangered long-lived perennial plant Narcissus pseudonarcissus using random amplified polymorphic markers. Estimates for mean gene diversity within 15 populations of N. pseudonarcissus of three neighbouring geographical regions were high in comparison to other long-lived perennials (H _eN = 0.33). The genetic diversity of the two smallest populations (<200 plants) was significantly reduced, indicating loss of genetic variability due to drift. The analysis of the population genetic structure revealed a significant genetic differentiation both between regions (Φ_ST = 0.06) and between populations within regions (Φ_ST = 0.20). However, there was incomplete correspondence between geographical regions and the population genetic structure. In order to preserve the overall genetic variation in wild populations of N. pseudonarcissus, management measures should thus aim to protect many populations in each region. The spatial genetic structure within populations of N. pseudonarcissus was in agreement with an isolation by distance model indicating limited gene flow due to pollinator behaviour and restricted seed dispersal. The very restricted spatial extent of clonal growth (<5 cm) and the high level of clonal diversity indicate that clonal growth in N. pseudonarcissus is not an important mode of propagation and that management measures should favour sexual reproduction in order to avoid further reductions in the size and number of populations.     

Cryptic changes in the genetic structure of a highly clonal coral population and the relationship with ecological performance

Elkhorn coral, Acropora palmata, relies heavily on clonal propagation and often displays low genotypic (clonal) diversity. Populations in the Florida Keys experienced rapid declines in tissue cover between 2004 and 2006, largely due to hurricanes and disease, but remained stable from 2006 to 2010. All elkhorn colonies in 150 m² permanent study plots were genotyped in 2006 (n = 15 plots) and 2010 (n = 24 plots), and plots sampled in both years were examined for changes in allelic and genotypic diversity during this period of stable ecological abundance. Overall, genetic diversity of Florida plots was low and declined further over the 4-yr period; seven of the 36 original genets and two of 67 alleles (among five microsatellite loci) were lost completely from the sampled population, and an additional 15 alleles were lost from individual reefs. In 2010, Florida plots (~19 colonies) contained an average of 2.2 ± 1.38 (mean ± SD) genets with a significant negative correlation between colony abundance and genotypic diversity. When scaled to total tissue abundance, genotypic diversity is even lower, with 43 % of genets below the size of sexual maturity. We examined the hypothesized positive relationship of local genotypic diversity with ecological performance measures. In Florida plots (n = 15), genotypic diversity was not significantly correlated with tissue loss associated with chronic predation, nor with acute disease and storm-fragmentation events, though this relationship may be obscured by the low range of observed diversity and potential confounding with abundance. When more diverse plots in Curaçao (n = 9) were examined, genotypic diversity was not significantly correlated with resistance during an acute storm disturbance or rate of recovery following disturbance. Cryptic loss of genetic diversity occurred in the apparently stable Florida population and confirms that stable or even increasing abundance does not necessarily indicate genetic stability.     

Genetic diversity of the endangered Chinese endemic herb Saruma henryi Oliv. (Aristolochiaceae) and its implications for conservation

Saruma henryi Oliv., the only representative of the monotypic genus Saruma Oliv. (Aristolochiaceae), is an endangered perennial herb endemic to China. It is a phylogenetically, ecologically, and medicinally important species. In the present study, inter-simple sequence repeat (ISSR) markers were employed to investigate the genetic diversity and differentiation of 14 populations. A total of 16 selected primers yielded 175 bright and discernible bands, with an average of 10.94 per primer. POPGENE analysis showed that the genetic diversity was quite low at the population level (h = 0.0447–0.1243; I = 0.0642–0.1853; PPB = 10.29–36.57%), but pretty high at the species level (h = 0.2603; I = 0.3857; PPB = 73.71%). The hierarchical analysis of molecular variance (AMOVA) revealed a high level of genetic differentiation among populations (67.18% of total variance components, P < 0.001), in line with the gene differentiation coefficient (G _ST = 0.6903) and the limited among-population gene flow (N _m = 0.2243). Both Principal Coordinates Analysis (PCoA) and UPGMA cluster analysis supported the grouping of all 14 populations into three geographic groups, among which there occurred a moderate level of genetic differentiation (33.18% of total variance components, P < 0.001) as shown by AMOVA analysis. In addition, Mantel test revealed a significant correlation between genetic and geographic distances among populations (r = 0.7792, P = 0.001), indicating the role of geographic isolation in shaping its present population genetic structure. The present levels and patterns of genetic diversity of S. henryi were assumed to result largely from its breeding system, geographic isolation, clonal growth, its unique biological traits and evolutionary history. The high genetic differentiation among populations implies that the conservation efforts should aim to preserve all the extant populations of this endangered herb.     

Conservation genetics in two endangered endemics from the Canary Islands, Helianthemum gonzalezferreri Marrero (Cistaceae) and Kunkeliella subsucculenta Kämmer (Santalaceae): different life histories that involve different management strategies

Helianthemum gonzalezferreri and Kunkeliella subsucculenta, two endangered endemic species from the Canary Islands, were analysed to determine the levels and structure of genetic diversity in their natural populations. The mean value of Shannon’s diversity index for H. gonzalezferreri and K. subsucculenta were I = 0.315 and I = 0.331, respectively. AMOVA and Bayesian analysis showed high genetic differentiation between populations in H. gonzalezferreri (F _ST = 0.297). This genetic differentiation was graphically shown with the principal component analysis (PCA), in which the majority of individuals were distributed into two groups. Contrarily, K. subsucculenta populations showed a considerable degree of gene exchange revealed by Bayesian cluster analysis and PCA. The genetic differentiation between the two H. gonzalezferreri populations suggests different management strategies for each population. Translocation between the two natural populations of K. subsucculenta, to increase genetic variation, would not mean outbreeding depression, since both populations share the same genetic pool.     

Quantifying the levels of sexual reproduction and clonal spread in the invasive plant, Rosa multiflora

Rosa multiflora Thunb., (Rosaceae), an invasive plant in the eastern U.S., was introduced into the U.S. in the early 1800s and was widely planted in the 1940s as a living fence, for wildlife cover, and to prevent soil erosion. This species spread rapidly from these original plantings via seed dispersal (sexual reproduction) and clonal spread, invading pasture and wooded areas. In this study we used allozyme markers to test for significant differences in the levels of asexual and sexual spread in large (>9 m cirumference) verses small (<2 m circumference) patches of R. multiflora and in pasture verses park settings. Although larger patches of R. multiflora tended to be dominated by one genotype, they exhibited significantly greater genetic diversity and inputs from sexual reproduction than did small patches; all large patches (N = 10) contained multiple unique genotypes. In contrast six of ten smaller patches of R. multiflora, consisted of a single genotype, though three patches had two genotypes and one had three unique genotypes. Similar analyses revealed clonal structure in R. multiflora populations both park and pasture habitats but with significantly greater genetic diversity and sexual inputs in the former than the latter. These results are consistent with a model of invasive spread involving clonal spread, sexual reproduction, and bird-mediated seed dispersal into established patches. Sexual inputs appear to be highest in larger patches and park habitats where perching sites for birds are most abundant. This flexible reproduction system likely contributes to the invasiveness of R. multiflora and to current management failures.     

Influences of the genetic neighborhood on ramet reproductive success in a clonal desert cactus

Clonal structure in clonal plants can affect sexual reproduction. Individual ramets can decrease reproduction if their neighbors are ramets of the same genet due to inbreeding depression or self-incompatibility. We assessed ramet reproductive success in the partial self-incompatible Ferocactus robustus (Cactaceae) as a function of floral display size in focal ramets and floral display size and clonal structure of their reproductive neighborhoods. Ramets were labeled, sized in number of stems, mapped and genetically identified through RAPD markers in one population. A pollen dispersal area of 15-m radius was established for each ramet to determine the clonal diversity in the neighborhoods. Flower production and fruit set were counted on a monthly basis during one reproductive season as a surrogate of ramet fitness. We expected a decrease in individual ramet reproductive success as a function of the number of reproductive ramets of the same genet in the neighborhood. A total of 272 sampled ramets revealed 116 multilocus genotypes, showing high clonal diversity in the population (G/N = 0.43, D = 0.98). Clonal diversity of neighborhoods ranged from 0.06 to 1 and fruit set varied from 0 to 76.9%. Individual ramet reproductive success was influenced by (1) mate availability, (2) floral display size of a genet within the reproductive neighborhood, and (3) the proportion of distinguishable genotypes. Floral display size of genets and ramets coupled with the genetic diversity within the reproductive neighborhood determines the low sexual reproduction in F. robustus.     

Analysis of population clonal diversity using microsatellite markers in the salt marsh sedge Carex scabrifolia in western Japan

Carex scabrifolia, a perennial herb that commonly grows on sandbars in lagoons and tidal estuaries, is threatened by habitat reduction in some areas of Japan. Clonal diversity and the extent of gene flow among ten populations located along the Seto Inland Sea and the Japan Sea in western Japan were examined using six microsatellite loci. From 299 samples, we detected 77 multilocus genotypes. The mean number of alleles per population was 2.8 and the mean clonal diversity was 0.23. Many populations consisted of small patches, and the mean number of genets per patch was 2.0. The average number of alleles per locus and clonal diversity were positively correlated with the number of patches within a watershed. Gene flow was detected between the Ichikawa River and the Ohta River populations along the Seto Inland Sea, and weak differentiation among populations located along the Seto Inland Sea was observed. Our results suggest that effective conservation of C. scabrifolia populations should include maintaining all patches within a watershed regardless of population size, thus promoting genotype preservation.     

The extent of clonality and genetic diversity in the rare Caldesia grandis (Alismataceae): Comparative results for RAPD and ISSR markers

Genetic variation and clonal diversity of three natural populations of the rare, highly clonal marsh herb Caldesia grandis Samuelsson were investigated using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. Both of the markers worked effectively in clone identification of C. grandis. RAPD markers detected more diversity than ISSR markers in the three populations examined. Of the 60 RAPD primers screened, seven produced highly reproducible bands. Using these primers, a total of 61 DNA fragments were generated with 52 (85.25%) being polymorphic indicating considerable genetic variation at the species level. Analysis of molecular variance (AMOVA) showed that a large proportion of genetic variation (81.5%) resided within populations, while only a small proportion (18.5%) resided among populations. With the use of 52 polymorphic RAPD markers, we were able to identify 127 genets among 342 samples from three populations. The proportion of distinguishable genets (PD: mean 0.37), Simpson's diversity index (D: mean 0.91), and evenness (E: mean 0.78) exhibited high levels of clonal diversity compared to other clonal plants. These results imply that sexual reproduction has played an important role at some time during the history of these populations. Nevertheless, the high level of diversity could have been also partially generated from somatic mutations, although this is unlikely to account for the high diversity generally found among C. grandis genets.     

Comparison of phenotypic and genetic clone delineation in quaking aspen, <Emphasis Type="Italic">Populus tremuloides</Emphasis>

Key message

Clonal delineation at nuclear microsatellites and phenotypic traits showed high correspondence and revealed an important role of both sexual and clonal reproduction for stand genetic structure.

Abstract

Quaking aspen (Populus tremuloides Michx.) grows throughout the northern and central portions of North America. Reproduction occurs both sexually via seeds and clonally from root suckers. Clonal delineation using morphological/phenological traits, and more recently, highly variable nuclear microsatellites have shown considerable variation in the size of clonal assemblies, and the relative importance of sexual versus clonal reproduction across the species range. In order to provide reliable estimates of genet size (N/G; ramets per sampled genet) and genotypic diversity (G/N; genets/ramets), and to compare genetic and phenotypic clone delineation, we characterized 181 sampled stems (ramets) at seven nuclear microsatellites, and morphological and phenological traits from six clones (genet size ≥11). Genotypic diversity was moderate (G/N = 0.18) and within the range reported in other studies across North America. Multivariate statistics revealed a high correspondence between genetic and phenotypic clone delineation, both with and without predefined genetic groups (94.2 %, 81.7 %). Moderate average genet size (5.6 ramets per genet) and the occurrence of genetically distinct single-ramet genets surrounded by larger genets suggested intermediate levels of sexual reproduction contributing to the genetic structure of this stand. Significant differences among genets were found for phenological and morphological traits such as bark thickness and leaf shape. However, most clones showed no significant differences in diameter growth which was likely caused by poor drainage in this high clay soil that inhibited the expression of genetic differences in growth.

     

Genetic variation and population differentiation of the endochitinase gene family in Pinus monticola

In the present study, we characterized nucleotide variations of the Pinus monticola class IV endochitinase (PmCh4) family. Using primers targeting at conserved amino acid motifs of plant class IV endochitinases, genomic DNA was amplified. Sequence data analysis identified five novel genes in the PmCh4 family with one pseudogene. Single nucleotide polymorphisms of the PmCh4 family were surveyed in seven open-pollinated seed lots representing diverse geographical distribution. Variable levels of average pairwise nucleotide diversity (π = 0.00422–0.02079) and relatively high levels of haplotype diversity (H _d = 0.85–0.96) were revealed at PmCh4 loci. Based on nucleotide variation, P. monticola populations were clustered into two main groups by phylogenetic analysis based on Nei’s genetic distance. The Mantel test revealed no correlation between geographical and genetic distances (r = ?0.11, P = 0.59). A further SNP genetic diversity study on more P. monticola populations throughout Western North America may help the design of sampling regimes for tree breeding, genetic conservation and assisted migration under climate change.     

Plasticity in fitness and fitness-related traits at ramet and genet levels in a tillering grass Panicum miliaceum under patchy soil nutrients

Seedling and root sprout recruitment after a volcanic deforestation was examined in a Prunus ssiori population. According to genotypes of five microsatellite (SSR) loci, 44 genets were identified among 188 ramets ≥ 5 cm DBH in a 2.3-ha plot in 2000. The genets formed mutually exclusive patches. Because only one repeat change in an SSR locus distinguished five genets from adjacent genets, there is a possibility that the five genets derived from somatic mutations as well as sexual reproduction. Thus, at least 39 genets originated from seedlings, and at least 144 ramets derived from root sprouts. The seedlings were recruited after the volcanic eruptions in 1739 and before 1972 because trees of 5 cm DBH were 28 years of age, estimated from annual rings. As the largest ramet recruited from a root sprout was estimated to be 94 years old, root sprout recruitment began between 1739 and 1906. Thus, the estimated minimum rates of recruitment from seedlings and root sprouts were 0.167 y^–1 and 0.618-2.182 y^–1, respectively. The clonal diversity of the P. ssiori population (Simpson's D = 0.92) was higher than that of other clonal plant populations. The inbreeding coefficient was significantly negative (Wright's F _IS = −0.069). These results suggest that the frequent seedling recruitment in an outbreeding system maintains the clonal diversity in the population. This revised version was published online in June 2006 with corrections to the Cover Date.     

AFLP and ISSR analysis reveals high genetic variation and inter-population differentiation in fragmented populations of the endangered Litsea szemaois (Lauraceae) from Southwest China

Litsea szemaois (Lauraceae) is an endemic and endangered species from the tropical rain forests of Xishuangbanna, southern Yunnan, SW China, but habitat fragmentation, especially exacerbated by rubber planting, has caused a decline in population size of the species. AFLP and ISSR were used to investigate the genetic diversity and population structure of eight populations from across its known distribution. Three AFLP and ten ISSR primer combinations produced a total of 203 and 77 unambiguous and repeatable bands respectively, of which 164 (80.8%) and 67 (87.0%) were polymorphic for the two markers. These two markers showed that Litsea szemaois exhibits comparatively high genetic diversity at species level (heterozygosity (hs) = 0.2109) relative to some other Lauraceae. Most of the genetic variation was partitioned within populations, but genetic differentiation between populations was significant and relatively high (Φ _st = 0.2420, θ^B = 0.1986) compared with other tropical plants. The genetic characteristics of L. szemaois may be related to its outbreeding system, insect pollination and fragmented distribution. Because L. szemaois is dioecious and slow to mature, ex situ conservation across its genetic diversity is unlikely to succeed, although seedlings grow well under cultivation. Thus, in situ conservation is very important for this endangered species, especially as only 133 adult individuals are known in the wild. In particular, the Nabanhe and Mandian populations should be given a high conservation priority due to their higher genetic diversity, larger population size and better field condition, but wider sampling is required across all populations to determine additional areas with significant genetic conservation value.     

Population genetic structure of the endangered Eastern Bristlebird, Dasyornis brachypterus; implications for conservation

For species that are habitat specialists or sedentary, population fragmentation may lead to genetic divergence between populations and reduced genetic diversity within populations, with frequent inbreeding. Hundreds of kilometres separate three geographical regions in which small populations of the endangered Eastern Bristlebird, Dasyornis brachypterus, a small, ground-dwelling passerine that occurs in fire-prone bushland in eastern Australia, are currently found. Here, we use mitochondrial and microsatellite DNA markers to: (i) assess the sub-specific taxonomy designated to northern range-edge, and central and southern range-edge D. brachypterus, respectively, and (ii) assess levels of standing genetic variation and the degree of genetic subdivision of remnant populations. The phylogenetic relationship among mtDNA haplotypes and their spatial distribution did not support the recognised subspecies boundaries. Populations in different regions were highly genetically differentiated, but in addition, the two largest, neighboring populations (located within the central region and separated by ~50 km) were moderately differentiated, and thus are likely closed to migration (microsatellites, F _ST = 0.06; mtDNA, F _ST = 0.12, ?? _ST = 0.08). Birds within these two populations were genotypically diverse and apparently randomly mating. A long-term plan for the conservation of D. brachypterus??s genetic diversity should consider individual populations as separate management units. Moreover, managers should avoid actively mixing birds from different populations or regions, to conserve the genetic integrity of local populations and avoid outbreeding depression, should further translocations be used as a recovery tool for this species.     

Clonal and genetic structure of two Mexican oaks: Quercus eduardii and Quercus potosina (Fagaceae)

Quercus eduardii and Q. potosina are dominant oak species in Sierra Fría, Aguascalientes, Mexico. These species have been exploited for multiple purposes since the 16th century. Both species produce clonal offspring through root suckering and acorns through sexual reproduction. To understand clonality for the implementation of the most adequate actions for the conservation of these species, we addressed the following questions: (a) what is the spatial clonal structure of both species? (b) How much clonal and genetic diversity is maintained in these species? Random Amplified Polymorphic DNAs (RAPDs) were used as molecular markers for these analyses. Genets of both species have few ramets and these grow close the parent tree. Autocorrelation analyses at the ramet level showed an aggregated distribution at short distances and a random spatial distribution at larger distances. Also, at the genet level the autocorrelation analyses showed a random distribution. Clonal diversity was high in both species (Q. eduardii: D=0.963, G/N=0.60; Q. potosina: D=0.985, G/N=0.65). Genetic diversity was high within populations (Q. eduardii: H _e =0.33±0.11; Q. potosina: H _e =0.35±0.11). Low levels of genetic differentiation among populations were observed (Q. eduardii ? _st =0.19, P < 0.002; Q. potosina ? _st =0.13, P < 0.002). Both species maintain high levels of clonal and genetic diversity, probably due to successful sexual reproduction, which allows gene flow among populations. Conservation and/or reforestation programs must include seed collections and germplasm banks. Due to the small genet size and the high clonal diversity of these species, seeds can be collected in any place in Sierra Fría, Aguascalientes.     

Factors Affecting Distribution of Earthworms in Kashmir Valley: A Multivariate Statistical Approach

Soil characteristics influence earthworm population dynamics, species distribution and community structure. According in the present study an attempt was made to determine the soil physiochemical factors influencing earthworms of Kashmir valley with a view to improve the soil productivity by enhancing earthworm diversity under different pedoecosystems. Data collection on 15 soil parameters from 20 earthworm inhabiting sites revealed significant variation within and among the sites in soil temperature (F_{23, 19} = 148.83, 9.71; P < 0.05), moisture (F_{23, 19} = 16.91, 46.20; P < 0.05), pH (F₁₉ = 47.21; P < 0.05), electrical conductivity (F_{23, 19} = 11.67, 87.13; P < 0.05), sodium (F_{23, 19} = 2.46, 211.25; P < 0.05), potassium (F₁₉ = 22.91; P < 0.05), calcium (F₁₉ = 15.90; P < 0.05), magnesium (F_{23, 19} = 1.76, 104.51; P < 0.05), organic carbon (F_{23, 19} = 64.60, 222.50; P < 0.05), organic nitrogen (F_{23, 19} = 4.59, 3.81; P < 0.05) and phosphorous (F_{23, 19} = 5.11, 137.87; P < 0.05). Aporrectodea caliginosa trapezoides and A. rosea rosea exhibited wide range of distribution whereas Octolasion cyaneum, A. c. trapezoides and A. parva showed restricted distribution. Hierarchical cluster analysis grouped 20 earthworm collection sites into three clusters—earthworm absent sites, low earthworm diversity sites and moderate earthworm diversity sites. Principal component analysis assisted from the data set of 20 sites, resulting into four latent factors accounting for 77.95 % of total variance, identified the factors affecting earthworm communities are mainly related to physical habitat factor, chemical factor, soil texture factor and growth factor, each accounting for 26.41, 20.16, 18.25 and 13.13 % of total variance respectively.     

Genetic structure of a population of the clonal grass Setaria incrassata

We studied the genetic structure of a population of the clonal grass Setaria incrassata using isozyme electrophoresis to determine the extent of clone and gene diversity within and between three spatially isolated Setaria patches. High clone diversity and an even distribution of genets, which covered less than 0.25 m² on average, indicated that patch formation was dominated by propagation from seed. Gene diversity was high within the population and there was little genetic differentiation between patches. High levels of heterozygosity and polymorphism and strongly negative fixation indices indicated extensive recombination through outbreeding, despite the low number of alleles per locus. The synergistic effects of vegetative and sexual propagation may have contributed towards the unexpectedly high genetic diversity of this population. Genetic diversity in clonal populations may be preserved in the genotypes of clones and may therefore be increased by even rare recombination events.