Leaf epidermal characteristics of Asiatic Buddleja L. under scanning electron microscope: Insights into chromosomal and taxonomic significance

The leaf epidermis of all 23 species from Asian Buddleja was investigated with scanning electron microscopy (SEM). Characteristics of trichome type, stomatal shape, cuticular membrane on adaxial and abaxial epidermis, inner margin of outer stomatal ledge and ornamentation of outer stomatal ledge are presented in this study. Among the Buddleja species, some distinct epidermal patterns were observed. These can be used to characterize and identify different sections among the Asian Buddleja species, viz. Alternifoliae, Curviflorae and Rectiflorae. In general, stomatal length of Buddleja increases with ploidy level. This character can be used to distinguish diploids from polyploids without preparation of caryograms. Based on this correlation Buddleja microstachya, B. subcapitata and B. bhutanica are characterized to be diploids, as concluded from their stomatal dimensions. Leaf epidermis characters can also be used to differentiate species pairs that are easily confused with each other, viz., B. asiatica and B. bhutanica, B. albiflora and B. nivea, B. candida and B. fallowiana, B. delavayi and B. forrestii, B. japonica and B. lindleyana, B. officinalis and B. paniculata.     

Differential threshold effects of habitat fragmentation on gene flow in two widespread species of bush crickets

Effects of habitat fragmentation on genetic diversity vary among species. This may be attributed to the interacting effects of species traits and landscape structure. While widely distributed and abundant species are often considered less susceptible to fragmentation, this may be different if they are small sized and show limited dispersal. Under intensive land use, habitat fragmentation may reach thresholds at which gene flow among populations of small-sized and dispersal-limited species becomes disrupted. Here, we studied the genetic diversity of two abundant and widespread bush crickets along a gradient of habitat fragmentation in an agricultural landscape. We applied traditional (G(ST), θ) and recently developed (G'ST', D) estimators of genetic differentiation on microsatellite data from each of twelve populations of the grassland species Metrioptera roeselii and the forest-edge species Pholidoptera griseoaptera to identify thresholds of habitat fragmentation below which genetic population structure is affected. Whereas the grassland species exhibited a uniform genetic structuring (G(ST) = 0.020-0.033; D = 0.085-0.149) along the whole fragmentation gradient, the forest-edge species' genetic differentiation increased significantly from D < 0.063 (G(ST) < 0.018) to D = 0.166 (G(ST) = 0.074), once the amount of suitable habitat dropped below a threshold of 20% and its proximity decreased substantially at the landscape scale. The influence of fragmentation on genetic differentiation was qualitatively unaffected by the choice of estimators of genetic differentiation but quantitatively underestimated by the traditional estimators. These results indicate that even for widespread species in modern agricultural landscapes fragmentation thresholds exist at which gene flow among suitable habitat patches becomes restricted.     

Optimization of soil physical and chemical conditions for the bioremediation of creosote-contaminated soil

Mispah type soil (FAO : Lithosol) contaminated with >250 000 mg kg^-1 creosote was collected from the yard of a creosote treatment plant. The soils carbon, nitrogen and phosphorus contents were determined. Due to creosote contamination, thecarbon content of the soil was found to be 130,000 mg C kg^-1. This concentration was found to greatly affect the nitrogen content (0.08%). The phosphorus content was less affected (4.5%). It was estimated that a nutrient amendment to bring the soil to a C : N 10 : 1 would be adequate to stimulate microbial growth and creosote degradation. The soil was amended with a range of C : N ratios below and above the estimated ratio. In one of the treatments, the phosphorus content was amended. Sterile and natural controls were also set up. The soil was incubated at 30 °C on a rotaryshaker at 150 rpm in the dark for six weeks. Water content was maintained at 70% field capacity. The lowest nitrogen supplementation (C : N = 25 : 1) was more effective in enhancing microbial growth (3.12E + 05) and creosote removal (68.7%) from the soil. Additional phosphorus was not very effective in enhancing the growth of microorganisms and removal of creosote. The highest nitrogen supplementation(C : N = 5 : 1) did not enhance microbial growth and creosote removal.A relationship between mass loss and creosote removal was also observed. Phenolics and lower molecular mass polycyclic aromatic hydrocarbons (PAHs) were observed to be more susceptible to microbial degradation than higher molecular mass compounds. Nutrient concentration, moisture content and pH were thus observed to play very significant roles in the utilization of creosote in soil. These results are being used for the development of a bioremediation technology for the remediation of creosote contaminated soils in a treatment plant in South Africa.     

Effects of irradiation on the reproductive ability of Zonitoides arboreus, a snail pest of orchid roots

The effect of irradiation on the reproductive ability of the orchid snail, Zonitoides arboreus, a serious pest of potted orchids in Hawaii, was investigated. Weekly egg production averaged between 0.8 and 1.9 over a 9–wk period for snails not exposed to irradiation, and egg hatch averaged 61%. In comparison to untreated controls, irradiation of snails at the lowest dose tested (34–37 Gy) reduced egg production and egg hatch by 63% and 94%, respectively over a 9–wk period. None of the snails treated with levels of irradiation ≥ 69 Gy produced viable eggs. This is the first study measuring the effect of ionising irradiation on a terrestrial snail or slug species using sterilising doses. Overall, the results show that the reproductive ability of this snail species is affected by irradiation in a similar manner as for Biomphalaria glabrata, an aquatic snail for which the effects of irradiation have been studied in detail.     

Cytotoxic lignans from Larrea tridentata

Six lignans, including the cyclolignan 3,4'-dihydroxy-3',4'-dimethoxy-6,7'-cyclolignan, were isolated from the flowering tops of Larrea tridentata. Additionally the flavanone, (S)-4',5-dihydroxy-7-methoxyflavanone, was isolated for the first time from L. tridentata or any member of the family Zygophyllaceae. All of the compounds were assessed for their growth inhibitory activity against human breast cancer, human colon cancer and human melanoma cell lines. The lignans had IC50 values of 5-60 microM with the linear butane-type lignans being the most potent, and it was found that colon cancer cells were the least sensitive cell type tested. The relative potency of linear butane type lignans against human breast cancer appears to correlate positively with the number of O-methyl groups present on the molecule.     

Monitoring bioremediation in creosote-contaminated soils using chemical analysis and toxicity tests

Three soils with a history of creosote contamination (designated NB, TI and AC) were treated in bench-scale microcosms using conditions (nutrient amendment, moisture content and temperature) which had promoted mineralization of ¹⁴C-pyrene in a preliminary study. Bioremediation was monitored using the solid-phase Microtox test, seed germination and earthworm survival assays, SOS-chromotest, Toxi-chromotest and a red blood cell (RBC) haemolysis assay. Contaminant concentrations in the AC soil did not change after 150 days. Polycyclic aromatic hydrocarbon (PAH) concentrations decreased in the NB soil, and toxicity decreased overall according to the earthworm, seed germination and Microtox tests. Although total petroleum hydrocarbons (TPHs) in the TI soil were reduced following treatment, results of the earthworm, seed germination, RBC and Microtox tests suggested an initial increase in toxicity indicating that toxic intermediary metabolites may have formed during biodegradation. Toxicity testing results did not always correlate with contaminant concentrations, nor were the trends indicated by each test consistent for any one soil. Each test demonstrated a different capacity to detect reductions in soil contamination. Journal of Industrial Microbiology & Biotechnology (2000) 24, 132–139. Received 14 June 1999/ Accepted in revised form 12 November 1999     

Direct heat injury of roots of woody swamp species

1. 1.Three month-old seedlings of Taxodium distichum, Nyssa aquatica, Cephalanthus occidentalis and cuttings of Salix nigra were acclimated to simulated natural swamp conditions (ambient temperature, saturated soil) and then used to assess direct high temperature injury of root tissue.

2. 2.Electrolyte leakage from excised root tissue exposed for 30 min to temperatures ranging from 30 to 66°C was used to assess cellular injury.

3. 3.The relationship between leakage and temperature was sigmoidal for each species.

4. 4.Inflection point temperatures on the response curves, ranged from 45.4 to 51.0°C, were species-specific, and indicated differences in thermal tolerance of root membranes.

5. 5.Root of C. occidentalis and N. aquatica were more heat tolerant than roots of T. distichum and S. nigra.

Long‐distance geneflow and habitat specificity of the rock‐dwelling coppertail skink,Ctenotus taeniolatus

Rock boulders or ‘bush‐rocks’ provide essential habitat for many organisms and there has been interest in rehabilitating areas denuded of rock with artificial substitutes. We examine whether the density and size of bush rock influences the density of the coppertail skink (Ctenotus taeniolatus). The success of habitat rehabilitation is contingent on dispersal of rock‐dwelling organisms into areas that have been remediated. To gauge the likelihood of this we characterize geneflow of coppertail skinks among discrete patches of rocky habitat associated with ridge tops. We genotyped 154 individuals from seven localities at six microsatellite DNA loci and from a subset of these individuals we obtained sequence data from the mitochondrial ND4 region. Our field survey established that lizard density was positively associated with the availability of suitably sized bush‐rock (P < 0.001), highlighting the importance of maintaining this habitat element, or replacing it where it has been lost. Despite the presence of habitat features that might be presumed as barriers to dispersal for coppertail skinks, such as intervening gullies and dense vegetation, our genetic data demonstrated high levels of geneflow among rocky ridge tops. Levels of partitioning estimated by global F_ST were significant but low for both microsatellite (F_ST = 0.020) and mitochondrial data (F_ST = 0.113). Spatial autocorrelation of genotypic similarity supports our conclusion of regular longer‐distance geneflow, and we infer lower levels of dispersal in juveniles than in adults. This study suggests that dispersal of coppertail skinks can be sufficient to naturally colonize areas of restored habitat.