Mit Tropfenmikrofluidik zu Hochgeschwindigkeits-Biotechnologie

In recent years, microfluidic technologies were introduced for massively parallel cultivation and screening approaches. Individual cells can easily be singularized, compartmentalized, and cultivated from mixed inocula using droplet microfluidics. The generation of millions of droplets in a high-throughput manner enables studying diverse samples and combining the evaluation of genetic and phenotypic variants. It is a powerful tool to explore and exploit natural metabolic diversity.

Perpendicular Cultivation for Improved In-Row Weed Control in Organic Peanut Production

Intensive cultivation in organic peanut is partially effective, but in-row weed control remains problematic. In an attempt to improve in-row weed control, irrigated trials were conducted from 2011 to 2013 near Ty Ty, GA to determine the feasibility of early-season cultivation perpendicular to row direction using a tine weeder when integrated with other weed-control practices. Combinations of perpendicular cultivation (cultivation perpendicular to row direction), parallel cultivation (cultivation in the same direction of the rows), and banded applications of herbicides derived from natural sources were compared. Perpendicular cultivation improved overall weed control and peanut yield (two years of three), but this benefit was independent of weed control from any form of parallel cultivation. Additionally, tractor tire tracks from perpendicular cultivation across the rows repeatedly crushed peanut seedlings. Parallel cultivation with the tine weeder was generally more effective than parallel cultivation with sweeps, particularly for southern crabgrass and Texas millet. Herbicides derived from natural products were inconsistent in controlling dicot weeds, ineffective in controlling annual grasses, and did not protect peanut yield from weed interference.

Organic cofactors in the metabolism of Dehalococcoides mccartyi strains

Dehalococcoides mccartyi strains are strictly anaerobic organisms specialized to grow with halogenated compounds as electron acceptor via a respiratory process. Their genomes are among the smallest known for free-living organisms, and the embedded gene set reflects their strong specialization. Here, we briefly review main characteristics of published Dehalococcoides genomes and show how genome information together with cultivation and biochemical experiments have contributed to our understanding of Dehalococcoides physiology and biochemistry. We extend this approach by the detailed analysis of cofactor metabolism in Dehalococcoides strain CBDB1. Dehalococcoides genomes were screened for encoded proteins annotated to contain or interact with organic cofactors, and the expression of these proteins was analysed by shotgun proteomics to shed light on cofactor requirements. In parallel, cultivation experiments testing for vitamin requirements showed that cyanocobalamin (vitamin B 12 ), thiamine and biotin were essential supplements and that cyanocobalamin could be substituted by dicyanocobinamide and dimethylbenzimidazole. Dehalococcoides genome analysis, detection of single enzymes by shotgun proteomics and inhibition studies confirmed the expression of the biosynthetic pathways for pyridoxal-5-phosphate, flavin nucleotides, folate, S -adenosylmethionine, pantothenate and nicotinic acids in strain CBDB1. Haem/cytochromes, quinones and lipoic acids were not necessary for cultivation or dechlorination activity and no biosynthetic pathways were identified in the genomes.

Use of Ichip for High-Throughput In Situ Cultivation of “Uncultivable” Microbial Species

ABSTRACT One of the oldest unresolved microbiological phenomena is why only a small fraction of the diverse microbiological population grows on artificial media. The “uncultivable” microbial majority arguably represents our planet's largest unexplored pool of biological and chemical novelty. Previously we showed that species from this pool could be grown inside diffusion chambers incubated in situ, likely because diffusion provides microorganisms with their naturally occurring growth factors. Here we utilize this approach and develop a novel high-throughput platform for parallel cultivation and isolation of previously uncultivated microbial species from a variety of environments. We have designed and tested an isolation chip (ichip) composed of several hundred miniature diffusion chambers, each inoculated with a single environmental cell. We show that microbial recovery in the ichip exceeds manyfold that afforded by standard cultivation, and the grown species are of significant phylogenetic novelty. The new method allows access to a large and diverse array of previously inaccessible microorganisms and is well suited for both fundamental and applied research.

Molecular Analysis of Surfactant-Driven Microbial Population Shifts in Hydrocarbon-Contaminated Soil

ABSTRACT We analyzed the impact of surfactant addition on hydrocarbon mineralization kinetics and the associated population shifts of hydrocarbon-degrading microorganisms in soil. A mixture of radiolabeled hexadecane and phenanthrene was added to batch soil vessels. Witconol SN70 (a nonionic, alcohol ethoxylate) was added in concentrations that bracketed the critical micelle concentration (CMC) in soil (CMC′) (determined to be 13 mg g−1). Addition of the surfactant at a concentration below the CMC′ (2 mg g−1) did not affect the mineralization rates of either hydrocarbon. However, when surfactant was added at a concentration approaching the CMC′ (10 mg g−1), hexadecane mineralization was delayed and phenanthrene mineralization was completely inhibited. Addition of surfactant at concentrations above the CMC′ (40 mg g−1) completely inhibited mineralization of both phenanthrene and hexadecane. Denaturing gradient gel electrophoresis of 16S rRNA gene segments showed that hydrocarbon amendment stimulatedRhodococcus and Nocardia populations that were displaced by Pseudomonas and Alcaligenespopulations at elevated surfactant levels. Parallel cultivation studies revealed that the Rhodococcus population can utilize hexadecane and that the Pseudomonas andAlcaligenes populations can utilize both Witconol SN70 and hexadecane for growth. The results suggest that surfactant applications necessary to achieve the CMC alter the microbial populations responsible for hydrocarbon mineralization.

Some Aspects of The Recent History of American Historiography

My title is already long. Yet it does not say all that it should. My proper subject is certain aspects of the writing of American history in the United States during the last three-quarters of a century. But in order to deal with that I must say much that applies with equal force to American historiography at large during that period. In so doing, I run the risk of leaving, a false impression. There is a danger that I may suggest that the events of the earlier years, where I am speaking of what is more general, had issue in the later in something much narrower than was in fact the case. But that which I leave out will be much more familiar to my audience than it is to me; and it may be sufficient if I say that when I turn away from the general subject of the writing of history in the United States to the more particular one of the writing of American history, I do so not unaware, and trusting my audience to be much more fully aware, of the parallel cultivation by American historians of other fields of historical investigation.