Microbial inoculation influences arbuscular mycorrhizal fungi community structure and nutrient dynamics in temperate tree restoration

Soil microbial communities have a profound influence on soil chemical processes and subsequently influence tree nutrition and growth. This study examined how the addition of a commercial inoculum or forest‐collected soils influenced nitrogen (N) and phosphorus (P) dynamics, soil microbial community structure, and growth in Liriodendron tulipifera and Prunus serotina tree saplings. Inoculation method was an important determinant of arbuscular mycorrhizal fungi (AMF) community structure in both species and altered soil N dynamics in Prunus and soil P dynamics in Liriodendron. Prunus saplings receiving whole forest soil transfers had a higher rhizosphere soil carbon/nitrogen ratio and ammonia content at the end of the first growing season when compared to unmanipulated control saplings. Inoculation with whole forest soil transfers resulted in increased inorganic phosphorus in Liriodendron rhizosphere soils. The number of AMF terminal restriction fragments was significantly greater in rhizosphere soils of Liriodendron saplings inoculated with whole forest soil transfers and Prunus saplings receiving either inoculum source than control saplings. Forest soil inoculation also increased AMF colonization and suppressed stem elongation in Liriodendron after 16 months; conversely, Prunus AMF colonization was unchanged and stem elongation was significantly greater when saplings were inoculated with whole forest soil transfers. Longer term monitoring of tree response to inoculation will be essential to assess whether early costs of AMF colonization may provide long‐term benefits. This study provides insight into how practitioners can use microbial inoculation to alter AMF community structure and functioning, subsequently influencing tree growth and nutrient cycling during the restoration of degraded lands.     

Hydroxylation and glycosylation of the four conserved lysine residues in the collagenous domain of adiponectin. Potential role in the modulation of its insulin-sensitizing activity

It has recently been shown that the fat-derived hormone adiponectin has the ability to decrease hyperglycemia and to reverse insulin resistance. However, bacterially produced full-length adiponectin is functionally inactive. Here, we show that endogenous adiponectin secreted by adipocytes is post-translationally modified into eight different isoforms, as shown by two-dimensional gel electrophoresis. Carbohydrate detection revealed that six of the adiponectin isoforms are glycosylated. The glycosylation sites were mapped to several lysines (residues 68, 71, 80, and 104) located in the collagenous domain of adiponectin, each having the surrounding motif of GXKGE(D). These four lysines were found to be hydroxylated and subsequently glycosylated. The glycosides attached to each of these four hydroxylated lysines are possibly glucosylgalactosyl groups. Functional analysis revealed that full-length adiponectin produced by mammalian cells is much more potent than bacterially generated adiponectin in enhancing the ability of subphysiological concentrations of insulin to inhibit gluconeogenesis in primary rat hepatocytes, whereas this insulin-sensitizing ability was significantly attenuated when the four glycosylated lysines were substituted with arginines. These results indicate that full-length adiponectin produced by mammalian cells is functionally active as an insulin sensitizer and that hydroxylation and glycosylation of the four lysines in the collagenous domain might contribute to this activity.     

32 species validation of a new Illumina paired-end approach for the development of microsatellites

Development and optimization of novel species-specific microsatellites, or simple sequence repeats (SSRs) remains an important step for studies in ecology, evolution, and behavior. Numerous approaches exist for identifying new SSRs that vary widely in terms of both time and cost investments. A recent approach of using paired-end Illumina sequence data in conjunction with the bioinformatics pipeline, PAL_FINDER, has the potential to substantially reduce the cost and labor investment while also improving efficiency. However, it does not appear that the approach has been widely adopted, perhaps due to concerns over its broad applicability across taxa. Therefore, to validate the utility of the approach we developed SSRs for 32 species representing 30 families, 25 orders, 11 classes, and six phyla and optimized SSRs for 13 of the species. Overall the IPE method worked extremely well and we identified 1000s of SSRs for all species (mean = 128,485), with 17% of loci being potentially amplifiable loci, and 25% of these met our most stringent criteria designed to that avoid SSRs associated with repetitive elements. Approximately 61% of screened primers yielded strong amplification of a single locus.     

The arthropod,but not the vertebrate host or its environment,dictates bacterial community composition of fleas and ticks

Bacterial community composition in blood-sucking arthropods can shift dramatically across time and space. We used 16S rRNA gene amplification and pyrosequencing to investigate the relative impact of vertebrate host-related, arthropod-related and environmental factors on bacterial community composition in fleas and ticks collected from rodents in southern Indiana (USA). Bacterial community composition was largely affected by arthropod identity, but not by the rodent host or environmental conditions. Specifically, the arthropod group (fleas vs ticks) determined the community composition of bacteria, where bacterial communities of ticks were less diverse and more dependent on arthropod traits—especially tick species and life stage—than bacterial communities of fleas. Our data suggest that both arthropod life histories and the presence of arthropod-specific endosymbionts may mask the effects of the vertebrate host and its environment.     

Live-cell Imaging and Quantitative Analysis of Embryonic Epithelial Cells in Xenopus laevis

Embryonic epithelial cells serve as an ideal model to study morphogenesis where multi-cellular tissues undergo changes in their geometry, such as changes in cell surface area and cell height, and where cells undergo mitosis and migrate. Furthermore, epithelial cells can also regulate morphogenetic movements in adjacent tissues¹. A traditional method to study epithelial cells and tissues involve chemical fixation and histological methods to determine cell morphology or localization of particular proteins of interest. These approaches continue to be useful and provide "snapshots" of cell shapes and tissue architecture, however, much remains to be understood about how cells acquire specific shapes, how various proteins move or localize to specific positions, and what paths cells follow toward their final differentiated fate. High resolution live imaging complements traditional methods and also allows more direct investigation into the dynamic cellular processes involved in the formation, maintenance, and morphogenesis of multicellular epithelial sheets. Here we demonstrate experimental methods from the isolation of animal cap tissues from Xenopus laevis embryos to confocal imaging of epithelial cells and simple measurement approaches that together can augment molecular and cellular studies of epithelial morphogenesis.Download video file.^{(77M, mp4)}     

Can geometry control the coordination behaviour of 2-hydroxy-1-naphthaldehyde-N(4)-phenylthiosemicarbazone? A study towards its origin

2-Hydroxy-1-naphthaldehyde-N(4)-phenylthiosemicarbazone [H₂-Nap-ptsc] (H₂L) was reacted with square planar complexes of the type [MCl₂(EPh₃)₂] (where M = Ni or Pd; E = P or As) and octahedral [RuHCl(CO)(PPh₃)₃]. Square planar complexes ([Ni(Nap-ptsc)PPh₃)]; [Pd(Nap-ptsc)PPh₃)]; [Pd(Nap-ptsc)AsPh₃)]) gave products with only ONS tri coordinated thiosemicarbazone whereas octahedral ruthenium complex gave two products, one with NS and the other with ONS coordination. It is interesting to note that NS bi coordination leads to the formation of more strained four member ring. All the new complexes have been characterised by analytical, spectral, crystallographic and electrochemical methods.     

Quantitative distribution of presumptive archaeal and bacterial nitrifiers in Monterey Bay and the North Pacific Subtropical Gyre

The recent isolation of the ammonia-oxidizing crenarchaeon Nitrosopumilus maritimus has expanded the known phylogenetic distribution of nitrifying phenotypes beyond the domain Bacteria. To further characterize nitrification in the marine environment and explore the potential crenarchaeal contribution to this process, we quantified putative nitrifying genes and phylotypes in picoplankton genomic libraries and environmental DNA samples from coastal and open ocean habitats. Betaproteobacteria ammonia monooxygenase subunit A (amoA) gene copy numbers were low or undetectable, in stark contrast to crenarchaeal amoA-like genes that were broadly distributed and reached up to 6 x 10(4) copies ml(-1). Unexpectedly, in the North Pacific Subtropical Gyre, a deeply branching crenarchaeal group related to a hot spring clade (pSL12) was at times abundant below the euphotic zone. Quantitative data suggested that the pSL12 relatives also contain archaeal amoA-like genes. In both coastal and open ocean habitats, close relatives of known nitrite-oxidizing Nitrospina species were well represented in genomic DNA libraries and quantitative PCR profiles. Planktonic Nitrospina depth distributions correlated with those of Crenarchaea. Overall, the data suggest that amoA-containing Crenarchaea are more phylogenetically diverse than previously reported. Additionally, distributional patterns of planktonic Crenarchaea and Nitrospina species suggest potential metabolic interactions between these groups in the ocean's water column.     

Control of Changes in Mitochondrial Activities during Aging of Potato Slices

Aging of slices of potato tuber (Solanum tuberosum L.) in an aerated liquid medium induces a number of changes in mitochondrial activities. A nonphosphorylative, cyanide-insensitive electron transport pathway (alternate pathway) is brought into operation. The rate of oxidation of exogenous NADH increases markedly and the efficiency of phosphorylation with this substrate remains the same as it is in mitochondria isolated from fresh tissue slices. On the contrary, the rates of oxidation of succinate and malate do not increase while lower phosphorylative efficiencies indicate that a fraction of their electrons reaches oxygen through the alternate pathway. Chloramphenicol, a specific inhibitor of the mitochondrial protein-synthesizing system, has no effect whatsoever on these events. However, cycloheximide, which acts on the corresponding cytoplasmic system, prevents both the development of the alternate pathway and the rise in the rate of oxidation of exogenous NADH. These effects are interpreted as showing a specific control of the cytoplasmic protein-synthesizing system on the changes in mitochondrial oxidations during aging.