Metal content of dandelion (Taraxacum officinale) leaves in relation to soil contamination and airborne particulate matter.

The global distribution of the common dandelion (Taraxacum officinale Weber, sensu lato; Asteraceae), along with its ability to tolerate a wide range of environmental conditions, make this 'species' a particularly attractive candidate to evaluate for its value as a biological monitor of environmental metal contamination. To examine the metal content of dandelion leaves in relation to environmental metal levels, the concentrations of eight metals (Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) were analyzed in leaf and soil samples collected at 29 sites in the mid-western United States differentially impacted by pollution. Sites were chosen primarily to cover a range of annual mean 24-h airborne particulate matter < or = 10 microm (hereafter, PM10) exposure, with PM10 levels varying from those found in isolated rural areas to levels typical of the most industrialized urban locations in the mid-western United States. A positive. significant correlation was detected between soil concentrations of each metal and measures of PM10 at a site, signifying that airborne particulate matter is a good indicator of soil metal contamination. Leaf concentrations of four of the eight metals (Cr, Mn, Pb and Zn) examined were found to increase significantly as the soil levels of these metals increased, but the percentage of the total variation explained by the relationship in these cases was generally low. This latter finding, along with the lack of a significant relationship between leaf and soil concentrations for the four other metals, indicate that the factors affecting metal absorption from the soil by dandelions are complex and that, aside from soil metal concentrations, other soil, plant and/or other environmental factors affect metal uptake. There was also no evidence that leaf metal concentrations were positively correlated with PM10. In addition, the concentrations of some metals (Cu, Fe, Mn, Pb and Zn) were significantly higher in leaves collected in the fall compared to those collected at the same sites in the spring. These findings suggest that dandelions may not be a particularly effective tool for quantifying levels of environmental metal contamination, at least on the scale of pollution typifying industrialized urban areas of the mid-western United States.

Genetic diversity of Typha latifolia (Typhaceae) and the impact of pollutants examined with tandem-repetitive DNA probes.

Genetic diversity at variable-number-tandem-repeat (VNTR) loci was examined in the common cattail, Typha latifolia (Typhaceae), using three synthetic DNA probes composed of tandemly repeated "core" sequences (GACA, GATA, and GCAC). The principal objectives of this investigation were to determine whether: (1) the previously reported almost complete lack of polymorphism at allozyme loci in this species was indicative of a reduced amount of genetic diversity at VNTR loci as well; (2) VNTR markers were informative about possible clonal propagation; and (3) significant differences in genetic structure of sampling sites were associated with differences in environmental levels of pollutants at those sites. Previously, widespread sampling across the eastern United States, surveying across ten allozyme loci, has detected only two genotypes, involving a difference at a single locus, among 104 populations. In this study, the amount of genetic diversity detected at VNTR loci: (1) among ramets (N = 40; 40 genotypes detected) collected at ∼8-km intervals along a 320-km transect; (2) among ramets (N = 220; 117 genotypes detected) from five study sites separated by 50-3000 m; and (3) even among ramets within each study site [N = 44 per site; from 13 to 34 genotypes detected per site (270 m(2))] exceeds that previously found in those more geographically widespread allozyme surveys. Among the 260 ramets analyzed here, the mean number of bands scored per individual was 48.61 (SD = 2.80). Mean genetic similarity among ramets collected along the 320-km transect was 0.91, which was within the range of mean genetic similarity within the five study sites (range: 0.89-0.95). Among the five study sites, 61% of the samples analyzed appeared to be clonal ramets, with up to 12 clones detected for 44 ramets sampled within a site. Clones grew intermingled and ranged up to 39 m in extent. Permutation tests of genetic similarity revealed significant genetic differentiation between each of the five study sites. Consistent with the previous allozyme studies, T. latifolia was characterized by extremely low genetic variation relative to levels of polymorphism detected at VNTR loci in other plant species. Estimated heterozygosity among ramets along the 320-km transect ranged from 0.11 to 0.13, while that within the five study sites ranged from 0.05 to 0.12. Estimates of F(st) (0.32-0.41) also indicated considerable genetic subdivision among these stands. Significantly higher genetic diversity was detected at the two study sites that chemistry and toxicity data indicate to be the most severely impacted by pollutants. Although this correlation does not establish cause and effect, the results of this study indicate that the analysis of genetic diversity at VNTR loci may be a useful tool for monitoring anthropogenic-induced changes in the genetic structure of natural populations of plants.

GELSTATS: a computer program for population genetics analyses using VNTR multilocus probe data.

GELSTATS, a computer program for population genetics analyses utilizing genetic markers revealed with variable number tandem repeat (VNTR) multilocus probes, is described and made available (both as C++ source code and as an executable DOS program). The program calculates several population genetics parameters, including: (i) individual and population band numbers; (ii) population bands exhibiting complete linkage (redundant examples of such bands can be removed in subsequent analyses); (iii) similarity (fraction of bands shared) between individuals and average similarity within and between designated groups; (iv) estimated probability that two individuals chosen at random will have identical band profiles; (v) heterozygosity estimates for designated groups; and (vi) Fst estimates. Nonparametric permutation methods are used to assess the significance of differences in both within- and between-group similarity. A jackknife test for heterozygosity differences between groups is also computed. Examples of GELSTATS analyses illustrate some features of the program.

Inheritance in turnip of variable-number tandem-repeat genetic markers revealed with synthetic repetitive DNA probes.

Oligomers (16-26 mers) composed of short, tandemly repeated DNA sequences (3-10 bases) were used individually with their complementary oligomer in separate polymerase chain reactions (PCRs) that extended the number of repeats to make 15 different PCR synthetic tandem-repeat (STR) probes. These PCR-STR probes were used to examine the inheritance of variable-number tandem-repeat (VNTR) genetic markers from two parent plants of turnip (Brassica rapa L.) to 20 offspring. Following HinfI digestion and PCR-STR probing of Southern blots, interpretable variable parental and offspring band profiles were found with 9 of the 15 probes used. Each of these nine probes produced a unique set of fragments, and no cases of different probes revealing the same fragment were detected. Seventy-nine parental fragments were found and, of these, 65% (51) appeared to be heterozygous in one or both parents, with 52% (41) appearing to be heterozygous in one of the parents exclusively. That these fragments are transmitted as though heterozygous in the parents implies that they are derived from the nuclear complement of the genome. Chi-square analyses of the transmission of markers are, in general, consistent with Mendelian expectations, although three non-parental bands were found accounting for approximately 0.5% of these transmitted bands. For the fragments heterozygous in one of the parents exclusively, seven alleles exhibited complete linkage in three groups, 12 alleles were incompletely linked in six groups, and four allelic groups involving 11 alleles were identified. PCR-STR probes are relatively rapid to generate and apply (no cloning, clone screening, or sequencing steps are required), and have been shown to reveal VNTR genetic markers in a wide variety of plant species. These results add to the list of studies showing that VNTR genetic markers (and in this case, markers revealed by PCR-STR probes) are transmitted for the greater part in a Mendelian fashion.

Population and species variation of minisatellite DNA in Plantago.

Three Plantago species were surveyed for within- and between-population variation at DNA sequences detected with the M13 minisatellite probe. The levels and patterns of variation detected by this probe correspond to those expected from the mating systems of the species. The highly-selfing species P. major has a relatively low variability of minisatellite sequences within populations and considerable differentiation between populations. The outcrossing species P. lanceolata exhibits higher minisatellite variability within populations and moderate differentiation between populations. P. coronopus, with a mixed mating system, has levels of variation intermediate between P. major and P. lanceolata. The levels of variation within and between populations corresponds, in general, to the levels of allozyme variation determined in an earlier study. Mating system and population structure appear to have a major influence on M13-detected fragment variation.

Genetic variation detected by use of the M13 "DNA fingerprint" probe in Malus, Prunus, and Rubus (Rosaceae).

Recently, "DNA fingerprints" have been reported in a wide array of organisms. We used the M13 repeat probe on several genera and species in the angiosperm family Rosaceae. Four apple cultivars could be differentiated when any one of five restriction enzymes was used to analyze minisatellite DNA. Similarly, four individual trees of Prunus serotina (black cherry) exhibited different "fingerprints" with each of four enyzmes. A total of 14 Rubus (blackberries and raspberries) plants representing four species were investigated with two enzymes. Extensive inter-and intraspecific variation was found. However, some closely growing plants had identical "fingerprints", probably due to their being derived through vegetative propagation.

M13 repeat probe detects DNA minisatellite-like sequences in gymnosperms and angiosperms.

Several kinds of minisatellite DNA, all of which are composed of low to moderately repetitive DNA, have been identified in tetrapod genomes. While the repeating oligonucleotide elements (subrepeats) of a given minisatellite are virtually identical, subrepeat nucleotide composition differs between different minisatellites. Several minisatellites have exhibited moderate to high levels of restriction length polymorphism in a number of tetrapods. Such hypervariable markers provide powerful tools for genetic analyses in several fields of biology. Minisatellite applications have been restricted to tetrapods, but here we demonstrate that one probe, the M13 repeat probe previously used to detect minisatellites in humans and bovines, also reveals minisatellite-bearing endonuclease fragments in gymnosperms and angiosperms. While the plant minisatellites appear to be somatically stable within an individual, they often vary within species in potentially useful ways. These results demonstrate that minisatellite-like families may be distributed over a wide taxonomic range in eukaryotes, opening the possibility of a commensurately wide utility of minisatellite probes in genetic analyses.