Beneficial interactions between micro-organisms and roots

Microbial activity in the rhizosphere can have positive and negative effects on plants. Some of the beneficial processes act by minimizing the negative effects or by modifying the cropping environment to enhance productivity. Processes that are considered here include the provision of nitrogen, phosphorus and iron to the plant, the biocontrol of diseases and deleterious organisms and the stabilization of soil structures. The use of genetic engineering techniques in studying these processes and generating novel strains which may enhance them is discussed, along with the possible consequences of the release of organisms into the environment.     

Positive interactions between nitrogen-fixing legumes and four different neighbouring species in a biodiversity experiment

The importance of facilitative processes due to the presence of nitrogen-fixing legumes in temperate grasslands is a contentious issue in biodiversity experiments. Despite a multitude of studies of fertilization effects of legumes on associated nonfixers in agricultural systems, we know little about the dynamics in more diverse systems. We hypothesised that the identity of target plant species (phytometers) and the diversity of neighbouring plant species would affect the magnitude of such positive species interactions. We therefore sampled aboveground tissues of phytometers planted into all plots of a grassland biodiversity–ecosystem functioning experiment and analysed their N concentrations, δ¹⁵N values and biomasses. The four phytometer species (Festuca pratensis, Plantago lanceolata, Knautia arvensis and Trifolium pratensis) each belonged to one of the four plant functional groups used in the experiment and allowed the effects of diversity on N dynamics in individual species to be assessed. We found significantly lower δ¹⁵N values and higher N concentrations and N contents (amount of N per plant) in phytometer species growing with legumes, indicating a facilitative role for legumes in these grassland ecosystems. Our data suggest that the main driving force behind these facilitative interactions in plots containing legumes was reduced competition for soil nitrate (“nitrate sparing”), with apparent N transfer playing a secondary role. Interestingly, species richness (and to a lesser extent functional group number) significantly decreased δ¹⁵N values, N concentrations and N content irrespective of any legume effect. Possible mechanisms behind this effect, such as increased N mineralisation and nitrate uptake in more diverse plots, now need further investigation. The magnitude of the positive interactions depended on the identity of the phytometer species. Evidence for increased N uptake in communities containing legumes was found in all three nonlegume phytometer species, with a subsequent strong increase in biomass in the grass F. pratensis across all diversity levels, and a lesser biomass gain in P. lanceolata and K. arvensis. In contrast, the legume phytometer species T. pratense was negatively affected when other legumes were present in their host communities across all diversity levels.     

Multilevel interactions between ethylene and auxin in Arabidopsis roots

Hormones play a central role in the coordination of internal developmental processes with environmental signals. Herein, a combination of physiological, genetic, cellular, and whole-genome expression profiling approaches has been employed to investigate the mechanisms of interaction between two key plant hormones: ethylene and auxin. Quantification of the morphological effects of ethylene and auxin in a variety of mutant backgrounds indicates that auxin biosynthesis, transport, signaling, and response are required for the ethylene-induced growth inhibition in roots but not in hypocotyls of dark-grown seedlings. Analysis of the activation of early auxin and ethylene responses at the cellular level, as well as of global changes in gene expression in the wild type versus auxin and ethylene mutants, suggests a simple mechanistic model for the interaction between these two hormones in roots, according to which ethylene and auxin can reciprocally regulate each other's biosyntheses, influence each other's response pathways, and/or act independently on the same target genes. This model not only implies existence of several levels of interaction but also provides a likely explanation for the strong ethylene response defects observed in auxin mutants.     

Cortical bone failure mechanisms during screw pullout

An experimental and computational study of screw pullout from cortical bone has been conducted. A novel modification of standard pullout tests providing real time image capture of damage mechanisms during screw pullout was developed. Pullout forces, measured using the novel test rig, have been validated against standard pullout tests. Pullout tests were conducted, considering osteon alignment, to investigate the effect of osteons aligned parallel to the axis of the orthopaedic screw (longitudinal pullout) as well as the effect of osteons aligned perpendicular to the axis of the screw (transverse pullout). Distinctive alternate failure mechanisms, for longitudinally and transversely orientated cortical bone during screw pullout, were uncovered. Vertical crack propagation, parallel to the axis of the screw, was observed for a longitudinal pullout. Horizontal crack propagation, perpendicular to the axis of the screw, was observed for a transverse pullout. Finite element simulation of screw pullout, incorporating material damage and crack propagation, was also performed. Simulations revealed that a homogenous material model for cortical bone predicts vertical crack propagation patterns for both longitudinal and transverse screw pullout. A bi-layered composite model representing cortical bone microstructure was developed. A unique set of material and damage properties was used for both transverse and longitudinal pullout simulations, with only layer orientations being changed. Simulations predicted: (i) higher pullout forces for transverse pullout; (ii) horizontal crack paths perpendicular to screw axis for transverse pullout, whereas vertical crack paths were computed for longitudinal pullout. Computed results agreed closely with experimental observations in terms of pullout force and crack propagation.     

Belowground volatiles facilitate interactions between plant roots and soil organisms

Many interactions between organisms are based on the emission and perception of volatiles. The principle of using volatile metabolites as communication signals for chemo-attractant or repellent for species-specific interactions or mediators for cell-to-cell recognition does not stop at an apparently unsuitable or inappropriate environment. These infochemicals do not only diffuse through the atmosphere to process their actions aboveground, but belowground volatile interactions are similarly complex. This review summarizes various eucaryotes (e.g., plant (roots), invertebrates, fungi) and procaryotes (e.g., rhizobacteria) which are involved in these volatile-mediated interactions. The soil volatiles cannot be neglected anymore, but have to be considered in the future as valuable infochemicals to understand the entire integrity of the ecosystems.     

Song-type matching between neighbouring song sparrows

In our study population, neighbouring song sparrows typically share two or more of their 6-10 song types. In an earlier experiment, we found that established neighbours typically reply to playback of neighbour-shared song with a different song they share with that neighbour ('repertoire matching'), rather than with the same song ('type matching') or with a nonshared song. In the present experiment, we considered the hypothesis that type matching is a threat or warning signal (Krebs et al. 1981, Animal Behaviour, 29, 918-923). We tested the specific prediction that a bird is more likely to type-match early in the breeding season when territory boundaries are new and still unstable, and more likely to repertoire-match later in the season, once those boundaries have become well established. Birds were played a shared song of a new neighbour once early (April) and again late (June) in the breeding season. As predicted, early in the season birds usually type-matched the playback (73% of the trials) but late in the season they type-matched only rarely (18%); birds never replied (early or late) with a nonshared song type. Copyright 2000 The Association for the Study of Animal Behaviour.     

Intracellular interactions between the pathways of carbohydrate oxidation and nitrate assimilation in plant roots

Using density gradient techniques we have shown that in addition to a location within the cytoplasm all the enzymes of the pentose phosphate pathway are also present within the plastids of apical cells of pea roots. The data are discussed in relation to the hypothesis that the pentose phosphate pathway provides the NADPH for nitrite assimilation, the enzymes of which pathway have previously been shown to be located within the plastids of apical cells of pea roots.     

Changing interactions between astrocytes and neurons during CNS maturation

The environments of the developing brain and injured adult brain differ in their abilities to support axonal growth. To determine if astrocytes contribute to this difference, neurons were plated onto astrocytes cultured from the neonatal rat cortex and from the injured adult brain. Two patterns of neurite growth were observed in these two astrocyte culture systems. Neurons contacting the neonatal astrocytes had neurites that were twice as long as those contacting the injured adult astrocytes. Furthermore, in cultures with neonatal astrocytes, neurites faithfully followed the astrocytic processes, maximizing their contact, while in cultures of injured adult astrocytes, the neurites had a tendency to cross the processes orthogonally, minimizing their interaction with the astrocytes. When neurons were grown suspended over either neonatal or injured adult astrocytes, no difference in neurite length or the pattern of neurite growth was observed, indicating that neurite growth was not differentially affected by soluble factors released from the two populations of astrocytes. The addition of fetal calf serum, which is known to contain protease inhibitors, did not alter neurite growth when compared to serum-free medium, suggesting that a substantial difference in protease activity does not account for the variations in neurite length observed. Based on these results, it appears that the molecular components of the external surface of injured adult astrocytes do not support neurite growth to the same extent as those found on neonatal astrocytes. The differing abilities of these two populations of cultured astrocytes to support neurite growth in culture may reflect a change in the functional role of these cells that occurs during the development of the central nervous system.     

Dynamic interactions between limb segments during planar arm movement

Movement of multiple segment limbs requires generation of appropriate joint torques which include terms arising from dynamic interactions among the moving segments as well as from such external forces as gravity. The interaction torques, arising from inertial, centripetal, and Coriolis forces, are not present for single joint movements. The significance of the individual interaction forces during reaching movements in a horizontal plane involving only the shoulder and elbow joints has been assessed for different movement paths and movement speeds. Trajectory formation strategies which simplify the dynamics computation are presented.     

Mechanical interactions between dendritic cells and T cells correlate with T cell responsiveness

Ag recognition is achieved through the communication across intercellular contacts between T cells and APCs such as dendritic cells (DC). Despite remarkable progress in delineating detailed molecular components at the intercellular contacts, little is known about the functional roles of physical cross-junctional adhesion between T and DC in shaping T cell responses. In addition, the mechanisms underlying sensitivity and specificity of Ag discrimination by T cells at intercellular contacts remain to be elucidated. In this study, we use single-cell force spectroscopy to probe the mechanical interactions between DC and T cells in response to stimulation with a panel of altered peptide ligands. The results show that intercellular interactions of DC-T cell conjugates exhibited different ranges of interaction forces in peptide-dependent manners that match the ability of the peptides to activate T cells. Elevated calcium mobilization and IL-2 secretion by T cells were only promoted in response to antigenic peptides that induce strong interaction forces, suggesting that mechanically stable DC-T cell contacts are crucial for driving T cell activation. Strong interactions were not solely dependent on cell-surface molecules such as TCRs and the adhesion molecule LFA-1, but were also controlled by cytoskeletal dynamics and the integrity of membrane lipid rafts. These data provide novel mechanical insights into the effect of Ag affinity on intercellular contacts that align with T cell responsiveness.     

Chitosanase and chitinase activities in tomato roots during interactions with arbuscular mycorrhizal fungi or Phytophthora parasitica

New chitosanase acidic isoforms have been shown in Glomus mosseae-colonized tomato roots and their induction, together with the previously described mycorrhiza-related chitinase isoform, has been further corroborated in plants colonized with another Glomus species (G. intraradices),as well as in tomato roots colonized in vitro by Giaspora rosea. The induction of these chitosanase isoforms appears as a specific response to the arbuscular mycorrhizal (AM) symbiosis, and does not correspond to unspecific defence mechanisms, since these isoforms were not induced by the pathogen Phytophthora parasitica. Analysis by isoelectrofocusing showed two closely migrating chitinase isoforms, specific to mycorrhizal plants colonized either with G. mosseae or G. intraradices, and their isoelectric points were estimated to be 4.5 and 4.7. The estimated molecular mass of chitosanases was 20 kDa, and after isoelectrofocusing, the chitosanase activities were detected along the acidic pH range (6.5-3.5). Constitutive and induced isoforms were also investigated during a time-course study. In some experiments, chitin and chitosan were embedded together as substrates in polyacrylamide gels with the aim of studying the capacity of some isoforms to display both chitinase and chitosanase activities. In extracts from plants colonized with either G. mosseae or G. intraradices, some constitutive chitinases and the previously described mycorrhiza-related chitinase isoform, appeared to display chitosanase activity, while this bifunctional character was not found for the chitinases from non-mycorrhizal tissue, nor in Phytophthora-infected plants. These results suggest some diversity in the chitinase activities concerning substrate specificity in mycorrhizal plants. The possible implications of these observations in the functioning of the symbiosis is discussed.Key words: Arbuscular mycorrhizas, chitinases, chitosanases, Phytophthora parasitica, tomato, Lycoperiscon esculentum.      

Mechanical correction of dynamometer moment for the effects of segment motion during isometric knee-extension tests

The purpose of this study was to determine the effect of dynamometer and joint axis misalignment on measured isometric knee-extension moments using inverse dynamics based on the actual joint kinematic information derived from the real-time X-ray video and to compare the errors when the moments were calculated using measurements from external anatomical surface markers or obtained from the isokinetic dynamometer. Six healthy males participated in this study. They performed isometric contractions at 90° and 20° of knee flexion, gradually increasing to maximum effort. For the calculation of the actual knee-joint moment and the joint moment relative to the knee-joint center, determined using the external marker, two free body diagrams were used of the Cybex arm and the lower leg segment system. In the first free body diagram, the mean center of the circular profiles of the femoral epicondyles was used as the knee-joint center, whereas in the second diagram, the joint center was assumed to coincide with the external marker. Then, the calculated knee-joint moments were compared with those measured by the dynamometer. The results indicate that 1) the actual knee-joint moment was different from the dynamometer recorded moment (difference ranged between 1.9% and 4.3%) and the moment calculated using the skin marker (difference ranged between 2.5% and 3%), and 2) during isometric knee extension, the internal knee angle changed significantly from rest to the maximum contraction state by about 19°. Therefore, these differences cannot be neglected if the moment-knee-joint angle relationship or the muscle mechanical properties, such as length-tension relationship, need to be determined.     

Molecular interactions between fibronectin and integrins during mouse blastocyst outgrowth

To investigate the mechanism of trophoblast adhesion to fibronectin, we cultured blastocysts in serum-free medium on proteolytic fibronectin fragments containing its major functional domains, and localized fibronectin-binding integrins in outgrowing trophoblast cells by immunofluorescent staining. Outgrowth comparable to that obtained with intact fibronectin was observed using a 120 kD chymotryptic fragment containing the central cell-binding domain (FN-120) and the Arg-Gly-Asp (RGD) recognition sequence. A 40 kD COOH-terminal chymotryptic fragment of fibronectin containing both a heparin-binding region and an alternate (non-RGD) cell-binding site was inactive in supporting trophoblast adhesion. Three synthetic peptides derived from the heparin-binding domain, including the CS1 alternate cell-binding site, were also unable to promote trophoblast cell adhesion. A 75 kD recombinant protein, ProNectin F, containing 13 copies of the cell recognition epitope of fibronectin, Val-Thr-Gly-Arg-Gly-Asp-Ser-Pro-Ala-Ser, vigorously supported blastocyst outgrowth. Blastocyst outgrowth was not significantly different when surfaces were precoated with cellular fibronectin, which contains an alternatively spliced type III repeat and is the form actually encountered in vivo. Several putative fibronectin receptors were localized in trophoblast outgrowths by immunofluorescent labeling. Antibodies reactive with integrin subunits α3, α5, αllb, αv, β1 and β3, but not α4, all bound to trophoblast cells. Antibodies raised against either the β1 or β3 integrin subunits significantly inhibited fibronectin-mediated outgrowth. These findings demonstrate the key role of the central cell-binding domain of fibronectin in trophoblast adhesion, and suggest four RGD-binding integrins, α3β1, α5β1, αllbβ3, and αvβ3, that could mediate trophoblast adhesion in vitro and may play an important role during implantation. © 1995 Wiley-Liss, Inc.     

Azoarcus spp. and their interactions with grass roots

The current knowledge on the divergence within the genus Azoarcus and about interactions with grasses is summarized. Grass-associated members of this genus of diazotrophs have only been isolated from a salt- and flood-tolerant pioneer plant in Pakistan, Kallar grass (Leptochloa fusca (L.) Kunth). Members of these bacteria belong to the beta subclass of the Proteobacteria, most closely related to purple bacteria such as Rhodocyclus purpureus. The isolates from one single host plant showed a surprising divergence, consisting of five groups of Azoarcus distinct at the species level. Molecular diagnostic tests, which are based on 16S ribosomal RNA sequences, allowed preliminary assignment of isolates to Azoarcus by PCR amplification and sequencing of PCR products. Moreover, the moleculer tests enabled us to detect an unculturable strain in Kallar grass roots, stressing that classical cultivation techniques at times fail to detect some groups of the microbial population. Using similar techniques, sequences rooting in the Azoarcus clade were also detected in field-grown rice, indicating that the natural host range might extend to rice. In gnotobiotic laboratory cultures, a member of Azoarcus is able to colonize rice roots endophytically: bacteria invade the roots in the zone of elongation and differentiation, colonize the cortex intra- and inter-cellularly, and penetrate deeply into the vascular system, entering xylem vessels, allowing systemic spreading into the rice shoot. Recently, we detected expression of nitrogenase of Azoarcus cells inside roots of rice seedlings, a result encouraging us to analyze interactions with rice in detail.     

The power of modified instrumental variables tests for unit roots

In this paper we investigate the power of Hall's (1989) instrumentalvariables tests for unit roots. A slight modification is madeto Hall's t statistic so that the modified t statistic can beapplied directly without encountering the possible negativestandard error problem of Hall's original t statistic. The majorfinding is that the power curves of the instrumental variablestests are not monotonic functions of the autoregressive coefficientof the unit root models when there is a negative moving averagecomponent.