Molybdenum-induced effects on leaf ultra-structure and rhizosphere phosphorus transformation in Triticum aestivum L.

Soil phosphorus (P) occurs in pools of lower availability due to soil P fixation and therefore, it is a key constrain to crop production. Long term molybdenum-induced effects in wheat and rhizosphere/non-rhizosphere soil P dynamics have not yet been investigated. Here, a long term field experiment was conducted to explore these effects in wheat consisting of two treatments i.e. with molybdenum (+Mo) and without molybdenum (-Mo). The results revealed that molybdenum (Mo) supply increased plant biomass, grain yield, P uptake, preserved the configuration of chloroplast, stomata, and mesophyll tissue cells, suggesting the complementary effects of Mo on wheat yield and P accumulation. During the periods of vegetative growth, soil organic carbon, organic matter, and microbial biomass P were higher and tended to decrease in rhizosphere soil at maturity stage. In +Mo treatment, the most available P fractions [H2O-Pi (16.2-22.9 mg/kg and 4.24-7.57 mg/kg) and NaHCO3-Pi (130-149 mg/kg and 77.2-88 mg/kg)] were significantly increased in rhizosphere and non-rhizosphere soils, respectively. In addition, the +Mo treatment significantly increased the acid phosphatase activity and the expression of phoN/phoC, aphA, olpA/lppC gene transcripts in rhizosphere soil compared to -Mo. Our research findings suggested that Mo application has increased P availability not only through biochemical and chemical changes in rhizosphere but also through P assimilation and induced effects in the leaf ultra-structures. So, it might be a strategy of long term Mo fertilizer supply to overcome the P scarcity in plants and rhizosphere soil.

Putative Neural Network Within an Olfactory Sensory Unit for Nestmate and Non-nestmate Discrimination in the Japanese Carpenter Ant: The Ultra-structures and Mathematical Simulation.

Ants are known to use a colony-specific blend of cuticular hydrocarbons (CHCs) as a pheromone to discriminate between nestmates and non-nestmates and the CHCs were sensed in the basiconic type of antennal sensilla (S. basiconica). To investigate the functional design of this type of antennal sensilla, we observed the ultra-structures at 2D and 3D in the Japanese carpenter ant, Camponotus japonicus, using a serial block-face scanning electron microscope (SBF-SEM), and conventional and high-voltage transmission electron microscopes. Based on the serial images of 352 cross sections of SBF-SEM, we reconstructed a 3D model of the sensillum revealing that each S. basiconica houses > 100 unbranched dendritic processes, which extend from the same number of olfactory receptor neurons (ORNs). The dendritic processes had characteristic beaded-structures and formed a twisted bundle within the sensillum. At the "beads," the cell membranes of the processes were closely adjacent in the interdigitated profiles, suggesting functional interactions via gap junctions (GJs). Immunohistochemistry with anti-innexin (invertebrate GJ protein) antisera revealed positive labeling in the antennae of C. japonicus. Innexin 3, one of the five antennal innexin subtypes, was detected as a dotted signal within the S. basiconica as a sensory organ for nestmate recognition. These morphological results suggest that ORNs form an electrical network via GJs between dendritic processes. We were unable to functionally certify the electric connections in an olfactory sensory unit comprising such multiple ORNs; however, with the aid of simulation of a mathematical model, we examined the putative function of this novel chemosensory information network, which possibly contributes to the distinct discrimination of colony-specific blends of CHCs or other odor detection.

Ultra-thin resin embedding method for scanning electron microscopy of individual cells on high and low aspect ratio 3D nanostructures.

The preparation of biological cells for either scanning or transmission electron microscopy requires a complex process of fixation, dehydration and drying. Critical point drying is commonly used for samples investigated with a scanning electron beam, whereas resin-infiltration is typically used for transmission electron microscopy. Critical point drying may cause cracks at the cellular surface and a sponge-like morphology of nondistinguishable intracellular compartments. Resin-infiltrated biological samples result in a solid block of resin, which can be further processed by mechanical sectioning, however that does not allow a top view examination of small cell-cell and cell-surface contacts. Here, we propose a method for removing resin excess on biological samples before effective polymerization. In this way the cells result to be embedded in an ultra-thin layer of epoxy resin. This novel method highlights in contrast to standard methods the imaging of individual cells not only on nanostructured planar surfaces but also on topologically challenging substrates with high aspect ratio three-dimensional features by scanning electron microscopy.