Nutrient limitation and botanical diversity in wetlands: can fertilisation raise species richness?

The 'resource balance hypothesis' proposes that the species richness of grassland vegetation is potentially highest when the N:P ratio of plant tissues is 10–15 (co-limitation), so that species richness could be raised by fertilisation with N or P at sites with lower or higher N:P ratios, respectively. Here we use data from field surveys in Swiss, Dutch and American fens or wet grasslands to analyse what changes in N:P ratios might produce noticeable changes in species richness. Plant species numbers, above-ground biomass, tissue N and P concentrations and soil pH were recorded in plots of 0.06–4 m². In each data set, plots with intermediate tissue N:P ratios (6–20) were on average most species-rich, but N:P ratios explained only 5–37% of the variation in species richness. Moreover, these effects were partially confounded with those of vegetation biomass and/or soil pH. The unique effects of N:P ratios (excluding those shared with biomass and pH) explained 11–17% of variation in species richness. The relationship between species richness and N:P ratios was asymmetric: plots with high N:P ratios were more species-poor than those with low N:P ratios. This was paralleled by a smaller species pool size at high N:P ratios (estimated from species numbers in multiple records), suggesting that fewer species are adapted to P-limited conditions than to N-limited conditions. According to these data, species richness in wetlands may possibly be raised by P-fertilisation when the initial N:P ratio of the vegetation is well above 20, but this option is not recommended for nature conservation as it might promote common species at the expense of rare ones.     

Isolation and characterization of heat-sensitive, thermotolerant defective mutants of CHO cells

A replica plating technique was utilized to isolate stable CHO cell mutants that are heat-sensitive and have altered capacities to develop thermotolerance. From a mutagen (EMS) treated population of CHO cells, two strains were isolated. One (HS-36) shows a greatly reduced ability to develop thermotolerance following an initial 45.0 degrees C heat shock. The other (HS-23) also shows a greatly reduced thermotolerance development following a short 45.0 degrees C induction dose, but a greater thermotolerance development following longer 45.0 degrees C induction doses. The dose-survival response following single-dose 45.0 degrees C heating of HS-23 cells suggests the presence of a resistant subpopulation which is not due to contamination from, or reversion to, wild-type cells. Both strains have unique morphological characteristics. Spheroids develop in the central portion of HS-36 colonies, though cells in monolayers are indistinguishable from wild-type parental cells. HS-23 cells grow in firmly attached monolayers, but more than 95% maintain a "rounded" morphology. The remainder show a "flattened" morphology typical of CHO cells. Both strains have parental CHO characteristics as determined by chromosome number, population doubling times, and survival responses to UV light and to gamma rays. Each has maintained its heat-sensitive and altered thermotolerance phenotype for a period of over 6 months in continuous log-phase culture.     

Wetland dicots and monocots differ in colonization by arbuscular mycorrhizal fungi and dark septate endophytes

As an initial step towards evaluating whether mycorrhizas influence composition and diversity in calcareous fen plant communities, we surveyed root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytic fungi (DSE) in 67 plant species in three different fens in central New York State (USA). We found colonization by AMF and DSE in most plant species at all three sites, with the type and extent of colonization differing between monocots and dicots. On average, AMF colonization was higher in dicots (58±3%, mean±SE) than in monocots (13±4%) but DSE colonization followed the opposite trend (24±3% in monocots and 9±1% in dicots). In sedges and cattails, two monocot families that are often abundant in fens and other wetlands, AMF colonization was usually very low (<10%) in five species and completely absent in seven others. However, DSE colonization in these species was frequently observed. Responses of wetland plants to AMF and DSE are poorly understood, but in the fen communities surveyed, dicots appear to be in a better position to respond to AMF than many of these more abundant monocots (e.g., sedges and cattails). In contrast, these monocots may be more likely to respond to DSE. Future work directed towards understanding the response of these wetland plants to AMF and DSE should provide insight into the roles these fungal symbionts play in influencing diversity in fen plant communities.     

Quantitative high-throughput measurement of gene expression with sub-zeptomole sensitivity by capillary electrophoresis

Microarray technologies have provided the ability to monitor the expression of whole genomes rapidly. However, concerns persist with regard to quantitation and reproducibility, and the detection limits for individual genes in particular arrays are generally unknown. This article describes a semiautomated PCR-based technology, Q-RAGE, which rapidly provides measurements of mRNA abundance with extremely high sensitivity using fluorescent detection of specific products separated by capillary electrophoresis. A linear relationship between template concentration and fluorescent signal can be demonstrated down to template concentrations in the low aM region, corresponding to approximately 0.04 zmol (24 molecules) per reaction. The technique is shown to be quantitative over five orders of magnitude of template concentration, and average mRNA abundances of approximately 0.01 molecule per cell can be detected. A single predefined set of 320 primers provides 90-95% coverage of all eukaryotic genomes. Analysis of a set of 19 p53-regulated genes in untreated cultures of normal human epithelial cells, derived from three different tissues, revealed a 600-fold range of apparent constitutive expression levels. For most of the genes assayed, good correlations were observed among the expression levels in normal mammary, bronchial, and epidermal epithelial cells.