Haptic Assistance Improves Tele-Manipulation With Two Asymmetric Slaves.

Tele-manipulation of heavy loads typically requires the simultaneous use of two asymmetric slaves: a crane for vertical weight support and a robot for accurate lateral positioning. The industrial standard prescribes a pair of operators for such tasks (one operator to control each slave), although in principle one operator might control both slaves with a single, hybrid interface. Accurate and safe co-operative handling of the expensive and fragile heavy components is difficult, presumably due to problems in the coordination of the subtasks and the lack of mutual awareness between the two operators. This study proposes a novel haptic assistance system to improve subtask coordination and task performance. Its novelty consists of haptically linking operators/interfaces through the joint task environment. The system's efficacy is evaluated with 15 pairs of co-operators and 15 individual uni-manual operators who maneuvered a heavy load through a bounded path in Virtual Reality. Haptic assistance improves task completion time for both groups. It also reduces control activity and self-reported workload without affecting a number of critical errors made by the operators. Moreover, without haptic assistance, uni-manual operators perform worse than co-operators, but this difference between the interfaces was not found with haptic assistance.

Tele-Manipulation with Two Asymmetric Slaves: Two Operators Perform Better Than One.

Certain tele-manipulation tasks require manipulation by two asymmetric slaves, for example, a crane for hoisting and a dexterous robotic arm for fine manipulation. It is unclear how to best design human-in-the-loop control over two asymmetric slaves. The goal of this paper is to quantitatively compare the standard approach of two co-operating operators that each control a single subtask, to a single operator performing bi-manual control over the two subtasks, and a uni-manual control approach. In a human factors experiment, participants performed a heavy load maneuvering and mounting task using a vertical crane and a robotic arm. We hypothesize that bi-manual control yields worse task performance and control activity compared to co-operation, because of conflicting spatial and temporal constraints. Literature suggests that uni-manual operators should perform better than co-operation, as co-operators critically depend on each other's actions. However, other literature provides evidence that individual operators have limited capabilities in controlling asymmetric axes of two dynamic systems. The results show that the two co-operators perform the maneuvering and mounting task faster than either bi- or uni-manual operators. Compared to co-operators, uni-manual operators required more control activity for the vertical crane and less for the robotic arm. In conclusion, this study suggests that when controlling two asymmetric slaves, a co-operating pair of operators performs better than a single operator.

Intrahaplotype polymorphism at the Brassica S locus.

The S locus receptor kinase and the S locus glycoproteins are encoded by genes located at the S locus, which controls the self-incompatibility response in Brassica. In class II self-incompatibility haplotypes, S locus glycoproteins can be encoded by two different genes, SLGA and SLGB. In this study, we analyzed the sequences of these genes in several independently isolated plants, all of which carry the same S haplotype (S(2)). Two groups of S(2) haplotypes could be distinguished depending on whether SRK was associated with SLGA or SLGB. Surprisingly, SRK alleles from the two groups could be distinguished at the sequence level, suggesting that recombination rarely occurs between haplotypes of the two groups. An analysis of the distribution of polymorphisms along the S domain of SRK showed that hypervariable domains I and II tend to be conserved within haplotypes but to be highly variable between haplotypes. This is consistent with these domains playing a role in the determination of haplotype specificity.

Haptic Shared Control in Tele-Manipulation: Effects of Inaccuracies in Guidance on Task Execution.

Haptic shared control is a promising approach to improve tele-manipulated task execution, by making safe and effective control actions tangible through guidance forces. In current research, these guidance forces are most often generated based on pre-generated, errorless models of the remote environment. Hence such guidance forces are exempt from the inaccuracies that can be expected in practical implementations. The goal of this research is to quantify the extent to which task execution is degraded by inaccuracies in the model on which haptic guidance forces are based. In a human-in-the-loop experiment, subjects (n = 14) performed a realistic tele-manipulated assembly task in a virtual environment. Operators were provided with various levels of haptic guidance, namely no haptic guidance (conventional tele-manipulation), haptic guidance without inaccuracies, and haptic guidance with translational inaccuracies (one large inaccuracy, in the order of magnitude of the task, and a second smaller inaccuracy). The quality of natural haptic feedback (i.e., haptic transparency) was varied between high and low to identify the operator's ability to detect and cope with inaccuracies in haptic guidance. The results indicate that haptic guidance is beneficial for task execution when no inaccuracies are present in the guidance. When inaccuracies are present, this may degrade task execution, depending on the magnitude and the direction of the inaccuracy. The effect of inaccuracies on overall task performance is dominated by effects found for the Constrained Translational Movement, due to its potential for jamming. No evidence was found that a higher quality of haptic transparency helps operators to detect and cope with inaccuracies in the haptic guidance.

Membrane proteins involved in pollen-pistil interactions.

Self-incompatibility (SI) is a widespread mechanism in angiosperms which prevents self-fertilization. This mechanism relies on cell-cell interactions between pollen and pistil. Among the different SI systems that have been reported, two have been particularly investigated: the gametophytic system of Solanaceae and the sporophytic system of Brassicaceae. In these two families, although the molecular bases of SI response are different, secreted and/or membrane-anchored proteins are required for self-pollen rejection. Interestingly, these proteins exhibit two functions: recognition and a catalytic activity. In this review article, we present recent advances which permit a better understanding of how these proteins control the male/female recognition event associated with the SI response.

A task-specific analysis of the benefit of haptic shared control during telemanipulation.

Telemanipulation allows human to perform operations in a remote environment, but performance and required time of tasks is negatively influenced when (haptic) feedback is limited. Improvement of transparency (reflected forces) is an important focus in literature, but despite significant progress, it is still imperfect, with many unresolved issues. An alternative approach to improve teleoperated tasks is presented in this study: Offering haptic shared control in which the operator is assisted by guiding forces applied at the master device. It is hypothesized that continuous intuitive interaction between operator and support system will improve required time and accuracy with less control effort, even for imperfect transparency. An experimental study was performed in a hard-contact task environment. The subjects were aided by the designed shared control to perform a simple bolt-spanner task using a planar three degree of freedom (DOF) teleoperator. Haptic shared control was compared to normal operation for three levels of transparency. The experimental results showed that haptic shared control improves task performance, control effort and operator cognitive workload for the overall bolt-spanner task, for all three transparency levels. Analyses per subtask showed that free air movement (FAM) benefits most from shared control in terms of time performance, and also shows improved accuracy.

Characterization of the gene encoding the plastid-located glutamine synthetase of Phaseolus vulgaris: regulation of beta-glucuronidase gene fusions in transgenic tobacco.

The gln-delta gene, which encodes the plastid-located glutamine synthetase of Phaseolus vulgaris, was cloned and its promoter region was sequenced. Primer extension analysis was used to map the two major transcription initiation sites which are about 90 nucleotides apart. A fusion of 2.3 kb of the upstream region of the gln-delta gene to the reporter gene uidA encoding beta-glucuronidase was shown to be expressed in the chlorophyllous cell types of leaves and stems and in the root meristem region of transgenic tobacco. Analysis of a series of three 5' promoter deletion fusions revealed the presence of a region essential for promoter activity between -786 and -327 and regions involved in tissue-specific regulation and light regulation between -786 and +43.

Rapid induction by wounding and bacterial infection of an S gene family receptor-like kinase gene in Brassica oleracea.

A receptor-like kinase, SRK, has been implicated in the autoincompatible response that leads to the rejection of self-pollen in Brassica plants. SRK is encoded by one member of a multigene family, which includes several receptor-like kinase genes with patterns of expression very different from that of SRK but of unknown function. Here, we report the characterization of a novel member of the Brassica S gene family, SFR2. RNA gel blot analysis demonstrated that SFR2 mRNA accumulated rapidly in response both to wounding and to infiltration with either of two bacteria: Xanthomonas campestris, a pathogen, and Escherichia coli, a saprophyte. SFR2 mRNA also accumulated rapidly after treatment with salicylic acid, a molecule that has been implicated in plant defense response signaling pathways. A SFR2 promoter and reporter gene fusion was introduced into tobacco and was shown to be induced by bacteria of another genus, Ralstonia (Pseudomonas) solanacearum. The accumulation of SFR2 mRNA in response to wounding and pathogen invasion is typical of a gene involved in the defense responses of the plant. The rapidity of SFR2 mRNA accumulation is consistent with SFR2 playing a role in the signal transduction pathway that leads to induction of plant defense proteins, such as pathogenesis-related proteins or enzymes of phenylpropanoid metabolism.

Natural antisense transcripts of the S locus receptor kinase gene and related sequences in Brassica oleracea.

Gene expression can be inhibited by antisense RNA transcripts. Although this phenomenon is widely used to analyse gene function in plants, the molecular mechanisms involved are poorly understood. One approach to improving our understanding of antisense gene regulation is to analyse the function of endogenous antisense transcripts. To date, only a small number of plant genes have been shown to be transcribed in both directions and limited information is available concerning the role of natural antisense transcripts in plants. In this study, we have identified several natural antisense transcripts which hybridise to probes derived from the S locus receptor kinase gene (SRK). The RNase protection assay and reverse trancriptase-PCR were used to demonstrate that a proportion of the antisense transcripts are encoded directly by SRK. Using different RNase protection probes, regions of the promoter, exon I (which encodes the S domain) and intron I of SRK were shown to be transcribed in an antisense direction. An antisense SRK transcript was shown to inhibit translation of a sense transcript in vitro. The possible role of antisense SRK transcripts in vivo is discussed.

The S-locus receptor kinase is inhibited by thioredoxins and activated by pollen coat proteins.

The self-incompatibility response in Brassica allows recognition and rejection of self-pollen by the stigmatic papillae. The transmembrane S-locus receptor kinase (SRK), a member of the receptor-like kinase superfamily in plants, mediates recognition of self-pollen on the female side, whereas the S-locus cysteine-rich protein (SCR) is the male component of the self-incompatibility response. SCR is presumably located in the pollen coat, and is thought to be the SRK ligand. Although many receptor-like kinases have been isolated in plants, the mechanisms of signal transduction mediated by these molecules remain largely unknown. Here we show that SRK is phosphorylated in vivo within one hour of self-pollination. We also show that, in vitro, autophosphorylation of SRK is prevented by the stigma thioredoxin THL1 in the absence of a ligand. This inhibition is released in a haplotype-specific manner by the addition of pollen coat proteins. Our data indicate that SRK is inhibited by thioredoxins and activated by pollen coat proteins.

Two large Arabidopsis thaliana gene families are homologous to the Brassica gene superfamily that encodes pollen coat proteins and the male component of the self-incompatibility response.

The male component of the self-incompatibility response in Brassica has recently been shown to be encoded by the S locus cysteine-rich gene (SCR). SCR is related, at the sequence level, to the pollen coat protein (PCP) gene family whose members encode small, cysteine-rich proteins located in the proteo-lipidic surface layer (tryphine) of Brassica pollen grains. Here we show that the Arabidopsis genome includes two large gene families with homology to SCR and to the PCP gene family, respectively. These genes are poorly predicted by gene-identification algorithms and, with few exceptions, have been missed in previous annotations. Based on sequence comparison and an analysis of the expression patterns of several members of each family, we discuss the possible functions of these genes. In particular, we consider the possibility that SCR-related genes in Arabidopsis may encode ligands for the S gene family of receptor-like kinases in this species.

Identification of a gene linked to the Brassica S (self-incompatibility) locus by differential display.

Self-incompatibility in Brassica is controlled by a complex locus, the S locus, that includes several expressed genes. Two S locus genes, SLG and SRK, are expressed in the stigma and have been implicated in self-pollen recognition. The male component of this recognition system is also predicted to be encoded by a gene at the S locus but this gene has not been identified to date. In this study, we have used differential display to screen for polymorphic, S-locus-linked genes that are expressed in anthers. This approach has allowed the identification of a gene, named S5J, which was shown to segregate completely with the S locus. We discuss the possible role of this gene in the self-incompatibility response and evaluate the utility of differential display for the identification of genes at specific genetic loci.

The integral membrane S-locus receptor kinase of Brassica has serine/threonine kinase activity in a membranous environment and spontaneously forms oligomers in planta.

To gain further insight into the mode of action of S-locus receptor kinase (SRK), a receptor-like kinase involved in the self-incompatibility response in Brassica, different recombinant SRK proteins have been expressed in a membranous environment using the insect cell/baculovirus system. Recombinant SRK proteins exhibited properties close to those of the endogenous stigmatic SRK protein and were found to autophosphorylate on serine and threonine residues in insect cell microsomes. Autophosphorylation was constitutive because it did not require the presence of pollen or stigma extracts in the phosphorylation buffer. Phosphorylation was shown to occur in trans, suggesting the existence of constitutive homooligomers of membrane-anchored recombinant SRK. To investigate the physiological relevance of these results, we have examined the oligomeric status of SRK in planta in cross-linking experiments and by velocity sedimentation on sucrose gradients. Our data strongly suggest that SRK is associated both with other SRK molecules and other stigma proteins in nonpollinated flowers. These findings may have important implications for our understanding of self-pollen signaling.

Expression of glutamine synthetase genes in roots and nodules of Phaseolus vulgaris following changes in the ammonium supply and infection with various Rhizobium mutants.

In this paper we have examined whether the four glutamine synthetase (gln) genes, expressed in roots and nodules of Phaseolus vulgaris are substrate-inducible by ammonium. Manipulation of the ammonium pool in roots, through addition and removal of exogenous ammonium, did not elicit any changes in the abundances of the four mRNAs thus suggesting that the gln genes in roots of this legume are neither substrate-inducible by ammonium nor derepressed during nitrogen starvation. In nodules the effect of the ammonium supply on expression of the gln genes has been examined by growing nodules under argon/oxygen atmospheres, or with a number of Fix- Rhizobium mutants, and following addition of exogenous ammonium. The results of these experiments suggest that the expression of the gln-gamma gene, which is strongly induced during nodule development, is primarily under a developmental control. However nitrogen fixation appears to have a quantitative effect on expression of gln-gamma as the abundance of this mRNA is about 2 to 4-fold higher under nitrogen-fixing conditions. This effect could not be mimicked by addition of exogenous ammonium and moreover is not specific to the gln-gamma gene as mRNA from a leghaemoglobin gene was similarly affected. Taken together these results have failed to find an effect of ammonium on specifically inducing the expression of glutamine synthetase genes in roots and nodules of P. vulgaris.

A nitrate reductase gene of the cyanobacterium Synechococcus PCC6301 inferred by heterologous hybridization, cloning and targeted mutagenesis.

DNA probes from the narG gene of Escherichia coli, which encodes the large polypeptide of respiratory nitrate reductase, show cross-hybridization at low stringency to a single region of the genome of the cyanobacterium Synechococcus PCC6301. This segment of cyanobacterial DNA was cloned as the insert of plasmid pDN1 and characterized. RNA complementary to pDN1 was shown to be substantially more abundant in nitrate grown cells of Synechococcus PCC6301 than in ammonium grown cells, thus parallelling the nitrate induction and ammonium repression of nitrate reductase activity in cultures of this cyanobacterium. A mutant of Synechococcus PCC6301 deficient in nitrate reductase activity was obtained after a potentially mutagenic transformation treatment using pDN1 as a donor. This mutant was restored to the wild type phenotype following stable integrative transformation with pDN1 DNA. Taken together these data suggest that pDN1 might encode a polypeptide of nitrate reductase. pDN1 is distinct from three clones of genes involved in nitrate assimilation that were isolated previously from the related cyanobacterium Synechococcus PCC7942 (Kuhlemeier et al., 1984a, J. Bact. 159, 36-41, and 1984b, Gene 31, 109-116).

The S locus receptor kinase gene encodes a soluble glycoprotein corresponding to the SKR extracellular domain in Brassica oleracea.

Self-incompatibility in Brassica is controlled by the S locus which contains at least two genes. SLG encodes a secreted S locus glycoprotein whilst SRK encodes a putative S locus receptor kinase which consists of three domains: an extracellular domain sharing extensive sequence identity with SLG, transmembrane region, and a cytoplasmic domain exhibiting a serine/threonine protein kinase activity. Here, the existence of truncated forms of the SRK protein corresponding to the extracellular domain of the putative receptor is reported. These proteins were detected by an antibody which recognizes the N-terminus of SRK3 and, in an F2 progeny segregating for the S3 haplotype, were only expressed in plants possessing the S3 haplotype. The truncated SRK proteins were expressed specifically in stigmas but, unlike the membrane-spanning SRK3 protein, were soluble and occurred as four different glycoforms sharing the same amino acid backbone as shown by deglycosylation experiments. Several SRK3 transcripts that may code for these truncated SRK3 proteins have been identified by RACE PCR, stigma cDNA library screening and RNA blot analysis. These transcripts are apparently generated by a combination of alternative splicing and the use of alternative polyadenylation signals. The existence of truncated forms of the S locus receptor kinase highlights some similarities between plant and animal receptor kinases. In animals, soluble extracellular domains of receptors have been described and, in some cases, have been shown to play a role in the modulation of signal transduction. By analogy, the soluble, truncated SRK proteins may play a similar role in the self-incompatibility response.

SLR3: a modified receptor kinase gene that has been adapted to encode a putative secreted glycoprotein similar to the S locus glycoprotein.

A new member of the S gene family, SLR3 (S-Locus Related 3), was identified in Brassica oleracea. This gene had a novel pattern of expression compared with previously described members of the family, being expressed in petals, sepals and vegetative apices, in addition to stigmas and anthers. Moreover, use of SLR3-derived probes in RNA blot and RACE-PCR (rapid amplification of cDNA ends-polymerase chain reaction) experiments has identified transcripts of genes closely related to SLR3 in leaves, cotyledons and, at high levels in developing anthers. SLR3 is not linked to the S locus but is linked to two or three closely related genes. Sequence analysis of the SLR3 gene indicates that it is derived from an ancestral receptor kinase gene that has been modified by a series of deletion events. As a result of these modifications, SLR3 is predicted to encode a secreted glycoprotein lacking both transmembrane and kinase domains. The putative SLR3 protein differs from the products of most other S gene family members in that several of the highly conserved cysteines have been lost. Within the S gene family, modification of receptor kinase genes by deletion may represent a general mechanism for the generation of genes encoding secreted glycoproteins.

A functional S locus anther gene is not required for the self-incompatibility response in Brassica oleracea.

The self-incompatibility (SI) response in Brassica involves recognition of self-pollen by the papillar cells of the stigma and is mediated by the products of genes localized at the S (self-incompatibility) locus. Two S locus genes, SRK and SLG, are thought to encode components of a receptor complex present in the female partner. The putative gene product of SLA, a third S locus-linked gene that is expressed specifically in anthers, is a candidate for the male component of the SI recognition system. The identification of a mutant SLA allele, interrupted by a large insert resembling a retrotransposon, in self-compatible Brassica napus initially suggested that SLA played an essential role in the SI response. In this study, we have characterized an SLA allele from a self-compatible B. oleracea var acephala line and show that it too is interrupted by a large insert. However, analysis of seven B. oleracea var botrytis lines exhibiting both self-compatible and self-incompatible phenotypes showed that these lines carry an S allele very similar or identical to that of the B. oleracea var acephala line and that the SLA gene is interrupted by an insert in all seven lines. The insertion of the putative retrotransposon was shown to interfere with gene expression, with no SLA transcripts being detected by RNA gel blot analysis in a self-incompatible B. oleracea var botrytis line carrying an interrupted SLA gene. These data indicate that a functional SLA gene is not required for the SI response in Brassica.

Further analysis of the interactions between the Brassica S receptor kinase and three interacting proteins (ARC1, THL1 and THL2) in the yeast two-hybrid system.

The yeast two-hybrid system was used to further characterize the interactions between the Brassica S receptor kinase (SRK) and three putative substrates, ARC1 and the two thioredoxin h proteins, THL1 and THL2. Interactions were generally detectable with kinase domains of both Class I and Class II SRKs. Chimeric constructs were made between the SRK910 kinase domain and the non-interacting Arabidopsis RLK5 kinase domain. Only one chimeric construct, SRR2, interacted with THL1 and THL2, while none of the chimeras were able to interact with ARC1. SRR2 is largely made up of RLK5 kinase domain with the N-terminal end being derived from the SRK910 kinase domain and was the only chimeric construct that retained kinase activity. Deletion or substitution of a conserved cysteine at the N-terminal end of the SRK910 kinase domain resulted in loss of interaction with THL1 and THL2, while the addition of this cysteine to a related receptor kinase, SFR1, conferred the ability to interact with the thioredoxin h proteins. In addition, substitution of the cysteines in the THL1 active site abolished the interaction. Lastly, the two Arabidopsis thioredoxin h clones most closely related to THL1 and THL2 were found to interact with the SRK kinase domains. Thus, the nature of the interaction of the thioredoxin h clones with SRK involves the reducing activity of these proteins and is restricted to the class of thioredoxin h proteins which have the variant CPPC active site.