Research Notes: Cotyledon Culture

A procedure for aseptically culturing immature soybean cotyledons has been developed to study the synthesis of seed storage proteins. Experiments were carried out so that one cotyledon from an embryo was compared to the second cotyledon. Cotyledons were normally incubated for 6 days at 25 C in light with gentle shaking

Linking Microbial Community Structure and Ecosystem Functions in Acidic Soil from Pennsylvania, USA

Microorganisms play a critical role in the structure and functioning of soil ecosystems. Within acidic soil across the northeastern United States and Canada, we have little understanding of the microbial diversity present and its relationship to the biochemical cycles. The current study is aimed at understanding the taxonomical and functional diversities in the acidic soil obtained from near various types of trees, how the diversities change as a function of depth, and the linkage between taxonomical and functional diversities. From eight sampling locations, soil samples were collected from three horizons (depths). The three depths were 0-10 cm (A), 11-25 cm (B), and 26-40 cm (C). Results indicate that across all the samples analyzed, Bradyrhizobium and Candidatus Solibacter are the most abundant bacteria in the soil microbiome. The differences in the soil microbiome across the samples were attributed to the abundance of individual organism’s present in the soil and not to the presence or absence of individual organisms. Subsystem level analysis of the soil microbiome sequences indicate that there is higher level of abundance of genes attributed to regulation and cell signaling. A low level of sequences were detected for sulfur metabolism, potassium metabolism, iron acquisition and metabolism, and phosphorous metabolism. Structure-functional analysis indicate that Bradyrhizobium, Rhodopseudomonas, and Burkholderia are the major organisms involved in the nutritional ecosystem functioning within acidic soil. Based on the results, we propose utilizing a consortium of these organisms as an environmentally friendly alternative to the use of chemicals to maintain soil fertility and ecosystem functioning

Raccoon Spatial Ecology in the Rural Southeastern United States

The movement ecology of raccoons varies widely across habitats with important implications for the management of zoonotic diseases such as rabies. However, the spatial ecology of raccoons remains poorly understood in many regions of the United States, particularly in the southeast. To better understand the spatial ecology of raccoons in the southeastern US, we investigated the role of sex, season, and habitat on monthly raccoon home range and core area sizes in three common rural habitats (bottomland hardwood, upland pine, and riparian forest) in South Carolina, USA. From 2018–2022, we obtained 264 monthly home ranges from 46 raccoons. Mean monthly 95% utilization distribution (UD) sizes ranged from 1.05 ± 0.48 km2 (breeding bottomland females) to 5.69 ± 3.37 km2 (fall riparian males) and mean monthly 60% UD sizes ranged from 0.25 ± 0.15 km2 (breeding bottomland females) to 1.59 ± 1.02 km2 (summer riparian males). Males maintained home range and core areas ~2–5 times larger than females in upland pine and riparian habitat throughout the year, whereas those of bottomland males were only larger than females during the breeding season. Home ranges and core areas of females did not vary across habitats, whereas male raccoons had home ranges and core areas ~2–3 times larger in upland pine and riparian compared to bottomland hardwood throughout much of the year. The home ranges of males in upland pine and riparian are among the largest recorded for raccoons in the United States. Such large and variable home ranges likely contribute to elevated risk of zoonotic disease spread by males in these habitats. These results can be used to inform disease mitigation strategies in the southeastern United States

LDRD final report on "fundamentals of synthetic conversion of CO2 to simple hydrocarbon fuels" (LDRD 113486).

Energy production is inextricably linked to national security and poses the danger of altering the environment in potentially catastrophic ways. There is no greater problem than sustainable energy production. Our purpose was to attack this problem by examining processes, technology, and science needed for recycling CO{sub 2} back into transportation fuels. This approach can be thought of as 'bio-inspired' as nature employs the same basic inputs, CO{sub 2}/energy/water, to produce biomass. We addressed two key deficiencies apparent in current efforts. First, a detailed process analysis comparing the potential for chemical and conventional engineering methods to provide a route for the conversion of CO{sub 2} and water to fuel has been completed. No apparent 'showstoppers' are apparent in the synthetic route. Opportunities to improve current processes have also been identified and examined. Second, we have also specifically addressed the fundamental science of the direct production of methanol from CO{sub 2} using H{sub 2} as a reductant

Multi-Messenger Gravitational Wave Searches with Pulsar Timing Arrays: Application to 3C66B Using the NANOGrav 11-year Data Set

When galaxies merge, the supermassive black holes in their centers may form binaries and, during the process of merger, emit low-frequency gravitational radiation in the process. In this paper we consider the galaxy 3C66B, which was used as the target of the first multi-messenger search for gravitational waves. Due to the observed periodicities present in the photometric and astrometric data of the source of the source, it has been theorized to contain a supermassive black hole binary. Its apparent 1.05-year orbital period would place the gravitational wave emission directly in the pulsar timing band. Since the first pulsar timing array study of 3C66B, revised models of the source have been published, and timing array sensitivities and techniques have improved dramatically. With these advances, we further constrain the chirp mass of the potential supermassive black hole binary in 3C66B to less than $(1.65\pm0.02) \times 10^9~{M_\odot}$ using data from the NANOGrav 11-year data set. This upper limit provides a factor of 1.6 improvement over previous limits, and a factor of 4.3 over the first search done. Nevertheless, the most recent orbital model for the source is still consistent with our limit from pulsar timing array data. In addition, we are able to quantify the improvement made by the inclusion of source properties gleaned from electromagnetic data to `blind' pulsar timing array searches. With these methods, it is apparent that it is not necessary to obtain exact a priori knowledge of the period of a binary to gain meaningful astrophysical inferences.Comment: 14 pages, 6 figures. Accepted by Ap

Cluster M Mycobacteriophages Bongo, PegLeg, and Rey with Unusually Large Repertoires of tRNA Isotopes

Genomic analysis of a large set of phages infecting the common hostMycobacterium smegmatis mc2155 shows that they span considerable genetic diversity. There are more than 20 distinct types that lack nucleotide similarity with each other, and there is considerable diversity within most of the groups. Three newly isolated temperate mycobacteriophages, Bongo, PegLeg, and Rey, constitute a new group (cluster M), with the closely related phages Bongo and PegLeg forming subcluster M1 and the more distantly related Rey forming subcluster M2. The cluster M mycobacteriophages have siphoviral morphologies with unusually long tails, are homoimmune, and have larger than average genomes (80.2 to 83.7 kbp). They exhibit a variety of features not previously described in other mycobacteriophages, including noncanonical genome architectures and several unusual sets of conserved repeated sequences suggesting novel regulatory systems for both transcription and translation. In addition to containing transfer-messenger RNA and RtcB-like RNA ligase genes, their genomes encode 21 to 24 tRNA genes encompassing complete or nearly complete sets of isotypes. We predict that these tRNAs are used in late lytic growth, likely compensating for the degradation or inadequacy of host tRNAs. They may represent a complete set of tRNAs necessary for late lytic growth, especially when taken together with the apparent lack of codons in the same late genes that correspond to tRNAs that the genomes of the phages do not obviously encode

Considerations and best practices in animal science 16S ribosomal RNA gene sequencing microbiome studies

Microbiome studies in animal science using 16S rRNA gene sequencing have become increasingly common in recent years as sequencing costs continue to fall and bioinformatic tools become more powerful and user-friendly. The combination of molecular biology, microbiology, microbial ecology, computer science, and bioinformatics—in addition to the traditional considerations when conducting an animal science study—makes microbiome studies sometimes intimidating due to the intersection of different fields. The objective of this review is to serve as a jumping-off point for those animal scientists less familiar with 16S rRNA gene sequencing and analyses and to bring up common issues and concerns that arise when planning an animal microbiome study from design through analysis. This review includes an overview of 16S rRNA gene sequencing, its advantages, and its limitations; experimental design considerations such as study design, sample size, sample pooling, and sample locations; wet lab considerations such as field handing, microbial cell lysis, low biomass samples, library preparation, and sequencing controls; and computational considerations such as identification of contamination, accounting for uneven sequencing depth, constructing diversity metrics, assigning taxonomy, differential abundance testing, and, finally, data availability. In addition to general considerations, we highlight some special considerations by species and sample type