Evaluation of deep root phenotyping techniques in tube rhizotrons

Selection for deep rooting is critical for the development of genotypes that are able to explore deep soil water and nutrients, particularly as agricultural resources become more limited. However, current root phenotyping techniques demand significant investments of time, money, and effort, and measurements on very young plants or plants grown under soilless culture. This study introduced four novel and simple techniques for fast evaluation of root depth in tube rhizotrons, which enable root observation around the transparent tube walls, and allow roots growing to natural size in semi-field conditions. The first and second innovations involve the introduction of ¹⁵N tracer and herbicide to the roots, which estimated root activity by measuring the responses of the shoots aboveground. The third involves placement of a cone deep in the rhizotron, to increase chances to observe more deep roots along the tube walls. The fourth involves measurement of roots that emerge from the rhizotron bottom. The reliability of these techniques were assessed in a series of five experiments during 2014 and 2015. These tests compared two pairs of genotypes that previous studies had shown to have mutually distinctive root traits: the spring wheat pair of ‘April bearded’ vs. ‘Dacke’; and the winter wheat pair of ‘Tabasco’ vs. ‘Genius’, with the first of each pair being the genotype known for deep rooting. Results showed that the new techniques were as good as or better than existing alternatives at accurately measuring root traits. In eight of the nine comparisons, the measurements were consistent with the expectations of root characteristics for these known genotypes. Importantly, the indirect root activity measures (herbicide and ¹⁵N) showed the same trend as the direct root observation techniques in all experiments, but higher ability to distinguish the genotypes and more promise for future upscaling for plant breeding.     

Development, characterization and diagnostic application of a monoclonal antibody specific for a proteinase K resistant Lawsonia intracellularis antigen

Proliferative enteropathy (PE) is one of the most important infections in pigs caused by Lawsonia intracellularis, an obligate intracellular bacterium. The purpose of the present investigation was to develop monoclonal antibodies with specificity to L. intracellularis useful both for diagnostic purposes (by immunohistochemistry) and for bacterial characterization. Several antibody producing hybridomas were established by fusion of mouse myeloma with spleen cells from BALB/c mice immunized with mucosa scrapings of the intestinal mucosa from a L. intracellularis infected pig. A monoclonal antibody (mAb), Law1-DK, isotyped as IgG2b was selected by indirect immunofluorescence antibody test (IFAT). Histological sections of the intestines from pigs affected by proliferative enteropathy and in vitro grown bacteria in cell culture were tested positive for the presence of L. intracellularis with the mAb. A molecule at 21 kDa was recognized by the mAb in a Western blotting analysis when a whole-cell preparation of L. intracellularis was run on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This antigen was released from L. intracellularis by mild heat treatment and was resistant to proteinase K digestion, suggesting it to be non-protein, e.g., lipopolysaccharide (LPS). This suggestion was supported by its presence in the aqueous phase of a phenol-water extract. The inhibitory effect of periodate oxidation on the antigen-antibody binding confirmed the participation of a carbohydrate epitope. The new mAb was tested highly specific for L. intracellularis by applying in situ hybridization with a L. intracellularis specific probe targeting 16S ribosomal RNA simultaneously with the IFAT.     

The influence of topography on the development of Alfisols on calcareous clayey till in Denmark

A detailed macro- and micro-morphological study of seven closely-adjacent soil profiles on calcareous clay tills in a long-established beech wood in central Denmark, shows that depth of decalcification, thickness of textural B horizon, development of umbric epipedons and development of albic sub-horizons are all related to slope inclination, slope form and to relative situation in the slope complex.The soils may be classified within four subgroups of Alfisols, while one is a Mollisol. The MAST of 8°C poses problems of typification at suborder level between frigid and mesic classes.Development of clay skins and of glaebules and concretions as indicators of the redeposition of mobilised components, are present to greater extent in stable slope situations; with least lateral subsoil water movement. Development of albic horizons is related to greater lateral water movement.Development of cutanic features by argilluviation - argillans - do not exclude features formed by redeposition of carbonates - calcitans. Soil plasmic fabrics with both features are termed calcisepic fabrics and prove deposition of clay and carbonates in the same soil horizons, usually the lower B and C(g) horizons.Morphologically the dark epipedons show intense humification and many fecal pellets because of the undisturbed and high plant productivity and intense biotic activity. The micromorphology is one of isotic argillasepic plasmic fabrics. The argillic horizons are dominantly insepic or vosepic, while the calcareous C horizons are either argillasepic, or calci-vosepic or calcisepic. It is possible that some of the carbonate reprecipitation post-dates the argilluviation, the carbonates derived during secondary dissolution in the suprajacent horizons.     

Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants

Summary Transgenic barley plants (Hordeum vulgare L. cv. Kymppi) were obtained by particle bombardment of various tissues. Immature embryos and microspore-derived cultures were bombarded with gold particles coated with plasmid DNA carrying the gene coding for neomycin phosphotransferase II (NPTII), together with plasmid DNA containing the gene for -glucuronidase (GUS).Bombarded immature embryos were grown to plants without selection and NPTII activity was screened in small plantlets. One plant proved to be transgenic (T₀). This chimeric plant passed the transferred nptII gene to its T₁ progeny. The presence of the nptII gene was demonstrated by the PCR technique and enzyme activity was analyzed by an NPTII gel assay. Four T₀ spikes and 15 T₁ offspring were transgenic. The integration and inheritance was confirmed by Southern blot hybridization. Transgenic T₂ and T₃ plants were produced by isolating embryos from green grains of transgenic T₁ and T₂ plants, respectively and growing them to plants. After selfing, the ratio of transgenic to non-transgenic T₂ offspring was shown to follow the rule of Mendelian inheritance. The general performance of transgenic plants was normal and no reduction in fertility was observed.Microspore-derived cultures were bombarded one and four weeks after microspore isolation. After bombardment, cultures were grown either with or without antibiotic selection (geneticin ^R or kanamycin). When cultures were grown without selection and regenerated plants were transferred to kanamycin selection in rooting phase, one out of a total of about 1500 plants survived. This plant both carried and expressed the transferred nptII gene. The integration was confirmed by Southern blot hybridization. This plant was not fertile.     

Characterisation of Non-Autoinducing Tropodithietic Acid (TDA) Production from Marine Sponge Pseudovibrio Species

The search for new antimicrobial compounds has gained added momentum in recent years, paralleled by the exponential rise in resistance to most known classes of current antibiotics. While modifications of existing drugs have brought some limited clinical success, there remains a critical need for new classes of antimicrobial compound to which key clinical pathogens will be naive. This has provided the context and impetus to marine biodiscovery programmes that seek to isolate and characterize new activities from the aquatic ecosystem. One new antibiotic to emerge from these initiatives is the antibacterial compound tropodithietic acid (TDA). The aim of this study was to provide insight into the bioactivity of and the factors governing the production of TDA in marine Pseudovibrio isolates from a collection of marine sponges. The TDA produced by these Pseudovibrio isolates exhibited potent antimicrobial activity against a broad spectrum of clinical pathogens, while TDA tolerance was frequent in non-TDA producing marine isolates. Comparative genomics analysis suggested a high degree of conservation among the tda biosynthetic clusters while expression studies revealed coordinated regulation of TDA synthesis upon transition from log to stationary phase growth, which was not induced by TDA itself or by the presence of the C10-acyl homoserine lactone quorum sensing signal molecule.     

The foot and ankle of Australopithecus sediba

A well-preserved and articulated partial foot and ankle of Australopithecus sediba, including an associated complete adult distal tibia, talus, and calcaneus, have been discovered at the Malapa site, South Africa, and reported in direct association with the female paratype Malapa Hominin 2. These fossils reveal a mosaic of primitive and derived features that are distinct from those seen in other hominins. The ankle (talocrural) joint is mostly humanlike in form and inferred function, and there is some evidence for a humanlike arch and Achilles tendon. However, Au. sediba is apelike in possessing a more gracile calcaneal body and a more robust medial malleolus than expected. These observations suggest, if present models of foot function are correct, that Au. sediba may have practiced a unique form of bipedalism and some degree of arboreality. Given the combination of features in the Au. sediba foot, as well as comparisons between Au. sediba and older hominins, homoplasy is implied in the acquisition of bipedal adaptations in the hominin foot.     

A partial pelvis of Australopithecus sediba

The fossil record of the hominin pelvis reflects important evolutionary changes in locomotion and parturition. The partial pelves of two individuals of Australopithecus sediba were reconstructed from previously reported finds and new material. These remains share some features with australopiths, such as large biacetabular diameter, small sacral and coxal joints, and long pubic rami. The specimens also share derived features with Homo, including more vertically oriented and sigmoid-shaped iliac blades, greater robusticity of the iliac body, sinusoidal anterior iliac borders, shortened ischia, and more superiorly oriented pubic rami. These derived features appear in a species with a small adult brain size, suggesting that the birthing of larger-brained babies was not driving the evolution of the pelvis at this time.     

Electronic monitoring in fisheries: Lessons from global experiences and future opportunities

Since the beginning of the 21st century, electronic monitoring (EM) has emerged as a cost‐efficient supplement to existing catch monitoring programmes in fisheries. An EM system consists of various activity sensors and cameras positioned on vessels to remotely record fishing activity and catches. The first objective of this review was to describe the state of play of EM in fisheries worldwide and to present the insights gained on this technology based on 100 EM trials and 12 fully implemented programmes. Despite its advantages, and its global use for monitoring, progresses in implementation in some important fishing regions are slow. Within this context, the second objective was to discuss more specifically the European experiences gained through 16 trials. Findings show that the three major benefits of EM were as follows: (a) cost‐efficiency, (b) the potential to provide more representative coverage of the fleet than any observer programme and (c) the enhanced registration of fishing activity and location. Electronic monitoring can incentivize better compliance and discard reduction, but the fishing managers and industry are often reluctant to its uptake. Improved understanding of the fisher's concerns, for example intrusion of privacy, liability and costs, and better exploration of EM benefits, for example increased traceability, sustainability claims and market access, may enhance implementation on a larger scale. In conclusion, EM as a monitoring tool embodies various solid strengths that are not diminished by its weaknesses. Electronic monitoring has the opportunity to be a powerful tool in the future monitoring of fisheries, particularly when integrated within existing monitoring programmes.