A centrifuge tube reactor for the determination of bacterial methane oxidation enrichment factors without influence of diffusion related isotope fractionation.

Biotransformation of methane at landfill sites can be estimated by applying compound specific stable isotope analysis of methane from the anaerobic and the cover layer surface zone. Next to these two input parameters, merely the knowledge of the carbon isotopic fractionation of the bacterial methane oxidation in terms of the enrichment factor (ε) is required. However, many factors and conditions have been described to affect ε. These include temperature, the applied landfill cover, the type of expressed methane monooxygenase (MMO), and cell density. In this work we investigated the microbial methane oxidation with respect to temperature and type of methanotrophic enrichment culture. A newly designed setup was used to overcome potential CH4-substrate limitations such as diffusion that could affect the determined values of ε by improper and inhomogeneous mixing. The isotopic fractionation was determined based on the stable carbon isotope analysis of methane and carbon dioxide. The obtained value for isotopic fractionation was ε22°C = -0.0136 ±â€¯0.0036. Also for the first time, bulk stable isotope analysis of bacterial cell mass was performed by flow injection analysis isotope ratio mass spectrometry.

Applying reverse stable isotope labeling analysis by mid-infrared laser spectroscopy to monitor BDOC in recycled wastewater.

Biological stability of treated wastewater is currently determined by methods such as biological oxygen demand, ATP-quantification, or flow-cytometric cell counting. However, the continuous increase in water reclamation for wastewater reuse requires new methods for quantifying degradation of biodegradable dissolved organic carbon (BDOC) ranging from very small to high concentrations of dissolved organic carbon (DOC). Furthermore, direct activity measures or absolute concentrations of BDOC are needed that produce comparable and reproducible results in all laboratories. Measuring carbon mineralization by CO2 evolution presents a suitable approach for directly measuring the microbial degradation activity. In this work, we investigated the extent of BDOC in water samples from effluent of a wastewater treatment plant and after purification by ultrafiltration over 204 days. BDOC monitoring was performed with the recently introduced reverse stable isotope labeling (RIL) analysis using mid-infrared spectroscopy for the monitoring of microbial CO2 production. Average BDOC degradation rates ranged from 0.11 to 0.32 mg L-1 d-1 for wastewater treatment plant effluent and from 0.03 to 0.22 mg L-1 d-1 after ultrafiltration. BDOC was degraded over >90 days indicating the long-term instability of the DOC. Degradation experiments over 88 days revealed first order kinetic rate constants for BDOC which corresponded to 12.7 ·â€¯10-3 d-1 for wastewater treatment plant effluent and 2.7 ·â€¯10-3 d-1 after ultrafiltration, respectively. A thorough sensitivity analysis of the RIL showed that the method is very accurate and sensitive with method detection limits down to 10 µg·â€¯L-1 of measured CO2.

Germinal reversion of an unstable mutation for anthocyanin pigmentation in soybean.

Plants of the "w4-mutable" line of soybean [Glycine max (L.) Merr.] are chimeral for anthocyanin pigmentation. Mutable plants produce both near-white and purple flowers, as well as flowers of mutable phenotype with purple sectors on near-white petals. It is established here that the mutable trait is conditioned by an unstable recessive allele of the w4 locus that conditions anthocyanin biosynthesis. The gene symbol w4-m is assigned to the mutable allele. Allele w4-m was derived from a stable, wild-type W4 progenitor allele and reverts at high frequency to a stable, wild-type W4 allele. Reversion occurs both early and late during the development of the germ line. Several experiments give estimates of germinal reversion frequency, indicating that approximately 6% of mutable alleles revert to wild-type from one generation to the next. Allele w4-m exhibits many features typical of an allele controlled by a transposable element.