Plant development in the absence of epiphytic microorganisms

Microorganisms (bacteria, fungi) are common residents of the roots, stems and leaves of higher plants. In order to explore the dependency of plant development on the presence of epiphytic microorganisms, the achenes (seeds) of sunflower (Helianthus annuus L.) were sterilized and germinated under aseptic conditions. The sterility of the seedlings was determined with the agar impression method. In seedlings from non-sterile seeds (control) that were likewise raised in a germ-free environment, all plant organs investigated (stem, cotyledons and primary leaves) were contaminated with bacteria. Hypocotyl elongation was not affected by epiphytic microorganisms. However, the growth rates of the cotyledons and primary leaves were higher in sterile seedlings compared with the control. The implications of this differential inhibition of organ development by epiphytic bacteria that are transmitted via the outer surface of the seed coat are discussed. We conclude that epiphytes in the above-ground phytosphere are not necessary for the development of the sunflower seedling.     

Influence of Methodology and Assumptions on Reported National Carbon Flux Inventories: An Illustration from the Canadian Forest Sector

The Intergovernmental Panel on Climate Change (IPCC) has developed guidelines to standardize the international reporting of greenhouse gas emissions and removals by signatory nations of the UN Framework Convention on Climate Change. With regard to forest sector carbon fluxes, the IPCC guidelines require only that those fluxes directly associated with human activities (i.e., harvesting and land-use change) be reported. In Canada, the Carbon Budget Model of the Canadian Forest Sector (CBM-CFS2) has been used to assess carbon fluxes from the entire forest sector. This model accounts for carbon fluxes associated with both anthropogenic and natural disturbances, such as wild fires and insects. We combined model results for the period 1985 to 1989 with additional data to compile seven different national carbon flux inventories for the forest sector. These inventories incorporate different system components under a variety of seemingly plausible assumptions, some of which are encouraged refinements to the default flux inventory described in the IPCC guidelines. The resulting estimated net carbon fluxes varied from a net removal of 185,000 kt carbon per year of the inventory period to a netemission of 89,000 kt carbon per year. Following the default procedures in the IPCC guidelines, while using the best available national data, produced an inventory with a net removal of atmospheric carbon. Adding the effect of natural disturbances to that inventory reversed the sign of the net flux resulting in a substantial emission. Including the carbon fluxes associated with root biomass in the first inventory increased the magnitude of the estimated net removal. The variability of these results emphasizes the need for a systems approach in constructing a flux inventory. We argue that the choice of which fluxes to include in the inventory should be based on the importance of these fluxes to the overall carbon budget and not on the perceived ease with which flux estimates can be obtained. The results of this analysis also illustrate two specific points. Even those Canadian forests which are most free from direct human interactions—forests in which no commercial harvesting occurs—are not in equilibrium, and their contribution to national carbon fluxes should be included in the reported flux inventory. Moreover, those forest areas that are subject to direct management are still substantially impacted by natural disturbances. The critical effect of inventory methodology and assumptions on inventory results has important ramifications for efforts to “monitor” and “verify” programs aimed at mitigating global carbon emissions.     

Prenatal cytogenetic diagnosis after transabdominal chorionic villus sampling in the first trimester

First trimester prenatal cytogenetic diagnosis was attempted in 350 pregnancies after trans-abdominal chorionic villus sampling. The cytogenetic investigation was performed using both a short-term method (24 h incubation) and cell culture. Adequate samples were obtained in 99·1 per cent and in all these cases the fetal karyotype was established. A chromosome abnormality was found in 2·0 per cent of cases. A discrepancy between the karyotype obtained after 24 h incubation and the karyotype in cell culture was observed in 2·3 per cent. Maternal cell contamination in the cultures was confirmed in 13 of 181 cases where the 24 h incubation revealed a male karyotype. Studies of culture morphology showed that colonies of convoluted cells may serve as a marker for contamination with maternal cells in culture. For the present, we recommend using a short-term method as well as cell culture for cytogenetic investigation until the problems with karyotype discrepancy and maternal cell contamination have been further clarified.     

A routine method for storing lymphocytes for repeated isolation of DNA

A protocol for easy storage and later expansion of lymphocyte populations is given. Compared with methods using transformed cell lines, the method has a number of advantages for repeated production of cells for the isolation of DNA in amounts sufficient for use in diagnostic DNA technology.