Zeitschriftenartikel zum Thema „Plant Mobile Domain (PMD)“

Silencing of transposable elements (TEs) is an essential process to maintain genomic integrity within the cell. InArabidopsis, together with canonical epigenetic pathways such as DNA methylation and modifications of histone tails, the plant mobile domain (PMD) proteins MAINTENANCE OF MERISTEMS (MAIN) and MAIN-LIKE 1 (MAIL1) are involved in TE silencing. In addition, the MICRORCHIDIA (MORC) ATPases, including MORC1, are important cellular factors repressing TEs. Here, we describe the genetic interaction and connection between the PMD and MORC pathways by showing thatMORC1expression is impaired inmainandmail1mutants. Transcriptomic analyses of higher order mutant plants combiningpmdandmorc1mutations, andpmdmutants in whichMORC1expression is restored, show that the silencing defects of a subset of TEs inpmdmutants are most likely the consequence ofMORC1down-regulation. Besides, a significant fraction of up-regulated TEs inpmdmutants are not targeted by the MORC1 pathway.

Abstract Association of RAG2 with the epigenetic marker H3K4me3 through a plant homeodomain (PHD) in the RAG2 non core domain is important for relieving auto inhibition of the RAG recombinase activity for V(D)J recombination. However, little is known regarding the properties of RAG2 that regulate its PHD-dependent interactions with H3K4me3. Accordingly, we measured the localization and dynamics of an N-terminal GFP fusion protein of full length RAG2 (GFP-FL); a T490A mutant of full length RAG2 (GFP-T490A) that does not undergo a regulatory phosphorylation proximal to the PHD region of RAG2; and GFP-labeled core domain of RAG2 (GFP-Core). The proteins were expressed in RAG2−/− pro-B cells, and the H3K4me3 was labeled using monoclonal antibody. Cross-correlation analysis showed that GFP-FL and GFP-Core exhibited similar colocalization with H3K4me3, which was significantly less than colocalization of GFP-T490A with H3K4me3. Blocking H3K4 demethylation using 2,4-pyridinedicarboxylic acid (PDA) caused GFP-FL/H3K4me3 colocalization to approach that of GFP-T490A and H3K4me3, whereas GFP-Core/H3K4me3 colocalization was unchanged. Super resolution imaging showed that RAG2 interactions with H3K4me3 were restricted to interfaces with H3K4me3-enriched puncta, and that GFP-T490A exhibited greater overlap with the H3K4me3-enriched puncta than GFP-FL. Finally, measurement of fluorescence recovery after photobleaching demonstrated that GFP-T490A had a lower mobile fraction and rate of diffusion than GFP-FL in conditions where RAG2 interactions with H3K4me3 were enhanced. These data suggest that RAG2 interactions with H3K4me3 are regulated by Thr490 proximal to the PHD region, such as by a phosphoryation/dephosphorylation cycle.

The torque load spectrum is an important basis for the strength design and durability test verification of tractor power take-off (PTO), and the performance and reliability of tractor PTO directly affect the quality and efficiency of agricultural operations. In this paper, taking the PTO torque load as the object, a PTO loading method based on the dynamic load spectrum acquired in the actual field work was proposed in this paper. Based on the Peak Over Threshold model, the extrapolation of the PTO load spectrum was realized, and the load spectrum throughout the whole life cycle was obtained. On the basis of this, the mobile tractor PTO loading test bench and Fuzzy-Proportional-Integral-Derivative (Fuzzy-PID) controller were developed to achieve the dynamic loading of the PTO load spectrum, and the dynamic characteristics were analyzed and verified by the simulation and laboratory test. The results showed that with the time domain extrapolation method, the load extreme value was expanded from (63.24, 469.50) to (60.88, 475.18), and the coverage was expanded by 1.98%. By comparing with the fitting results, statistical characteristics and rain flow counting results, the load spectrum extrapolation method was effective. In addition, the response time of simulation and laboratory test were 0.05s and 0.75s, respectively; the maximum error was 1.77% and 4.03%, respectively; and the goodness of fit was 16.78 N·m, which indicated that the PTO loading test bench, can accurately restore the dynamic loading of the tractor and the Fuzzy-PID controller had better accuracy and stability. It would provide a reference for the practical application of PTO load spectrum of the tractors.

Chromosomal translocations are genetic rearrangements where a chromosomal segment is transferred to a non-homologous chromosome which give rise to novel chimeras. Chromosomal rearrangements play a significant role in the development of acute leukemias (acute lymphoblastic leukemia (ALL) or acute myeloid leukemia (AML)). Chromosomal translocation events occurring at 11q23 involving the KMT2A or Mixed-Lineage Leukemia (MLL) gene (n=102) can be diagnosed in about 5-10% of all acute leukemia patients (Marschalek Ann Lab Med 2016), especially prevalent in infant acute leukemias (up to 70% of cases). Different chromosomal translocation partner genes (such as AF4, AF6, AF9orENL and ELL) account for the majority of leukemia cases and have their genomic breakpoints within a major breakpoint cluster region (BCR intron 9-11; Meyer et. al. Leukemia 2018). Some rearrangements are specifically associated with particular disease phenotype e.g. the majority of ALL patients (~ 90%) are mainly caused by the following gene fusions, MLL-AF4, MLL-AF9, MLL-ENL. We are interested in a rare but yet drastic chromosomal translocation t(6;11)(q27;q23) which fuses KMT2A/MLL to Afadin (AFDN/AF6) gene. This chromosomal rearrangement has a very poor prognosis (survival-rate is ~10%) and is predominantly diagnosed in patients with high-risk AML. In this project, we investigate the molecular consequences of two different MLL-AF6 fusions and their corresponding reciprocal AF6-MLL fusions. MLL-AF6 fusions are mainly occurring within MLL intron 9 to 11 and are associated with an AML disease phenotype, while the same fusion occurring within the minor breakpoints region in MLL intron 21 until exon (ex) 24 are mainly diagnosed with T-ALL (T-cell acute lymphoblastic leukemia) disease phenotype. The molecular mechanism that determines the resulting disease phenotype is yet unknown. Therefore, we cloned all of these t(6;11) fusion proteins in order to investigate the functional consequences of the two different breakpoints (MLLex1-9::AF6ex2-30, AF6ex1::MLLex10-37; MLLex1-21::AF6ex2-30, AF6ex1::MLLex22-37). All 4 fusion genes were introduced into our inducible Sleeping Beauty system (Ivics et. al. Mobile DNA 2010; Kowarz et. al. Biotechnol J. 2015) and stably transfected reporter cell lines. Basically, these 4 fusion proteins differ only in the presence or absence of their Plant homeodomain 1-3/Bromodomain (PHD1-3/BD) domain (see Figure 1). The PHD domain regulates the epigenetic and transcriptional regulatory functions of wildtype MLL. Subsequently, we analyzed gene expression differences by the MACE-Seq (Massive Analyses of cDNA Ends). MACE data revealed fundamental differences in gene expression profiles when analyzing the two different sets of t(6;11) fusion genes. The resulting profiles have similarities to either AML or T-ALL and might give a rational explanation for the different lineages in these t(6;11) patients. Altogether, these results notably indicate that our study will provide a novel insight into this type of high-risk leukemia and subsequently will be useful for developing of novel and appropriate therapeutic strategies against acute leukemia. Disclosures No relevant conflicts of interest to declare.

Ventilation systems are critically important components of many public buildings and workspaces. Proper ventilation is often crucial for preventing accidents, such as explosions in mines and avoiding health issues, for example, through long-term exposure to harmful respirable matter. Validation and maintenance of ventilation systems is thus of key interest for plant operators and authorities. However, methods for ventilation characterization, which allow us to monitor whether the ventilation system in place works as desired, hardly exist. This article addresses the critical challenge of ventilation characterization—measuring and modelling air flow at micro-scales—that is, creating a high-resolution model of wind speed and direction from airflow measurements. Models of the near-surface micro-scale flow fields are not only useful for ventilation characterization, but they also provide critical information for planning energy-efficient paths for aerial robots and many applications in mobile robot olfaction. In this article we propose a heterogeneous measurement system composed of static, continuously sampling sensing nodes, complemented by localized measurements, collected during occasional sensing missions with a mobile robot. We introduce a novel, data-driven, multi-domain airflow modelling algorithm that estimates (1) fields of posterior distributions over wind direction and speed (“ventilation maps”, spatial domain); (2) sets of ventilation calendars that capture the evolution of important airflow characteristics at measurement positions (temporal domain); and (3) a frequency domain analysis that can reveal periodic changes of airflow in the environment. The ventilation map and the ventilation calendars make use of an improved estimation pipeline that incorporates a wind sensor model and a transition model to better filter out sporadic, noisy airflow changes. These sudden changes may originate from turbulence or irregular activity in the surveyed environment and can, therefore, disturb modelling of the relevant airflow patterns. We tested the proposed multi-domain airflow modelling approach with simulated data and with experiments in a semi-controlled environment and present results that verify the accuracy of our approach and its sensitivity to different turbulence levels and other disturbances. Finally, we deployed the proposed system in two different real-world industrial environments (foundry halls) with different ventilation regimes for three weeks during full operation. Since airflow ground truth cannot be obtained, we present a qualitative discussion of the generated airflow models with plant operators, who concluded that the computed models accurately depicted the expected airflow patterns and are useful to understand how pollutants spread in the work environment. This analysis may then provide the basis for decisions about corrective actions to avoid long-term exposure of workers to harmful respirable matter.

Chaperonins cpn60 and cpn10 are essential proteins involved in cellular protein folding. Plant chloroplasts contain a unique version of the cpn10 co-chaperonin, cpn20, which consists of two homologous cpn10-like domains (N-cpn20 and C-cpn20) that are connected by a short linker region. Although cpn20 seems to function like other single domain cpn10 oligomers, the structure and specific functions of the domains are not understood. We mutated amino acids in the “mobile loop” regions of N-cpn20, C-cpn20 or both: a highly conserved glycine, which was shown to be important for flexibility of the mobile loop, and a leucine residue shown to be involved in binding of co-chaperonin to chaperonin. The mutant proteins were purified and their oligomeric structure validated by gel filtration, native gel electrophoresis, and circular dichroism. Functional assays of protein refolding and inhibition of GroEL ATPase both showed (i) mutation of the conserved glycine reduced the activity of cpn20, whether in N-cpn20 (G32A) or C-cpn20 (G130A). The same mutation in the bacterial cpn10 (GroES G24A) had no effect on activity. (ii) Mutations in the highly conserved leucine of N-cpn20 (L35A) and in the corresponding L27A of GroES resulted in inactive protein. (iii) In contrast, mutant L133A, in which the conserved leucine of C-cpn20 was altered, retained 55% activity. We conclude that the structure of cpn20 is much more sensitive to alterations in the mobile loop than is the structure of GroES. Moreover, only N-cpn20 is necessary for activity of cpn20. However, full and efficient functioning requires both domains.

There has been an increased emphasis on plant-based foods and diets. Although mobile technology has the potential to be a convenient and innovative tool to help consumers adhere to dietary guidelines, little is known about the content and quality of free, popular mobile health (mHealth) plant-based diet apps. The objective of the study was to assess the content and quality of free, popular mHealth apps supporting plant-based diets for Canadians. Free mHealth apps with high user ratings, a high number of user ratings, available on both Apple App and GooglePlay stores, and primarily marketed to help users follow plant-based diet were included. Using pre-defined search terms, Apple App and GooglePlay App stores were searched on December 22, 2020; the top 100 returns for each search term were screened for eligibility. Included apps were downloaded and assessed for quality by three dietitians/nutrition research assistants using the Mobile App Rating Scale (MARS) and the App Quality Evaluation (AQEL) scale. Of the 998 apps screened, 16 apps (mean user ratings±SEM: 4.6±0.1) met the eligibility criteria, comprising 10 recipe managers and meal planners, 2 food scanners, 2 community builders, 1 restaurant identifier, and 1 sustainability assessor. All included apps targeted the general population and focused on changing behaviors using education (15 apps), skills training (9 apps), and/or goal setting (4 apps). Although MARS (scale: 1–5) revealed overall adequate app quality scores (3.8±0.1), domain-specific assessments revealed high functionality (4.0±0.1) and aesthetic (4.0±0.2), but low credibility scores (2.4±0.1). The AQEL (scale: 0–10) revealed overall low score in support of knowledge acquisition (4.5±0.4) and adequate scores in other nutrition-focused domains (6.1–7.6). Despite a variety of free plant-based apps available with different focuses to help Canadians follow plant-based diets, our findings suggest a need for increased credibility and additional resources to complement the low support of knowledge acquisition among currently available plant-based apps. This research received no specific grant from any funding agency.

Throughout history, the quest for sustenance has been a constant struggle. With the advent of agriculture, humankind has witnessed remarkable progress over the centuries. In the present era, the Internet of Things (IoT) has permeated every sphere of life. Inspired by this, We have embarkedon a project to develop an IoT-based application tailored specifically for the agricultural domain, with a focus on revolutionizing plant management. Plant management encompasses a range of agricultural activities, including seedbed preparation, seed sowing, and plant maintenance. Ourwork aims to automate crucial processes such as water irrigation, water storage management, artificial lighting, and plant security systems. To achieve this, We have devised a dedicated mobile application that interfaces withIoT devices. These interconnected devices form a network, enabling real-time data monitoring and seamless activation of various operations within the system. This ambitious work consists of four major modules. To handle each module effectively, an individual nodemcu device has been allocated. By combining the power of IoT technology and agriculture, this work strives to optimize plant management, enhance productivity, and ultimately contribute to the advancement of this vital field

Abstract Helitrons are mobile elements found in the genomes of numerous plant and animal species. Helitrons can mobilize flanking sequences when they transpose, and so have the potential to duplicate genes and remodel genomes. The stony coral Acropora has an expanded immune gene repertoire in comparison to the closely-related anemone species Nematostella, especially in the number of TLR and IL-1R genes. Mechanisms behind the expansion are not currently known. Although helitron sequences have been identified in Nematostella, no helitrons have been identified in Acropora. BLAST searches against the Acropora genome using Nematostella amino acid sequences as queries revealed seven candidate Acropora helitrons. Six of the seven Acropora candidates had large open reading frames in the forward direction. One candidate element was split between two adjacent reading frames. Conserved domain searches were performed within the open reading frames using the databases at NCBI. As expected, Helitron N-like and replicative protein A (RPA) domains were shared between Acropora and Nematostella elements. UPGMA trees were constructed using highly conserved regions, and one candidate Acropora element clustered more closely with the Nematostella reference helitron than with other Acropora sequences. Interestingly, P-loop domains were common to cnidarian helitrons and a subset of plant helitron reference sequences containing plant disease resistance gene motifs in regions overlapping with a nucleotide-binding domain critical for resistance gene function. Together, these studies will help to determine if helitrons played a role in the expansion of the Acropora immune gene repertoire.

In today’s world, agricultural products are becoming increasingly scarce globally due to a variety of factors, and the early and accurate automatic identification of plant diseases can help ensure the stability and sustainability of agricultural production, improve the quality and safety of agricultural products, and help promote agricultural modernization and sustainable development. For this purpose, a lightweight deep isotropic convolutional neural network model, FoldNet, is designed for plant disease identification in this study. The model improves the architecture of residual neural networks by first folding the chain of the same blocks and then connecting these blocks with jump connections of different distances. Such a design allows the neural network to explore a larger receptive domain, enhancing its multiscale representation capability, increasing the direct propagation of information throughout the network, and improving the performance of the neural network. The FoldNet model achieved a recognition accuracy of 99.84% on the laboratory dataset PlantVillage using only 685k parameters and a recognition accuracy of 90.49% on the realistic scene dataset FGVC8 using only 516k parameters, which is competitive with other state-of-the-art models. In addition, as far as we know, our model is the first model that has fewer than 1M parameters while achieving state-of-the-art accuracy in plant disease identification. This proposal facilitates precision agriculture applications on mobile, low-end terminals.

Purpose – To avoid harm to humans, environment, and capital goods, hazardous or explosive gases that are possibly escaping from industrial and infrastructure facilities of the gas and oil processing industry have to be detected and located quickly and reliably. Project RoboGasInspector aims at the development and evaluation of a human-robot system that applies autonomous robots equipped with remote gas detection devices to detect and locate gas leaks. This article aims to focus on the usability of telemanipulation in this context. Design/methodology/approach – This paper presents four user studies concerning human-robot interfaces for teleoperation in industrial inspection tasks. Their purpose is to resolve contradictory scientific findings regarding aspects of teleoperation and to verify functionality, usability, and technology acceptance of the designed solution in the actual context of use. Therefore, aspects concerning teleoperation that were separately examined before are evaluated in an integrated way. Considered aspects are influence of media technology on telepresence, simulator sickness and head slaved camera control, usability of different input devices for telemanipulation, and identification of intuitive gestures for teleoperation of mobile robots. Findings – In general, the implemented interaction concepts perform better compared to conventional ones used in contemporary, actually applied robot systems. Otherwise, reasons are analyzed and approaches for further improvements are discussed. Exemplary results are given for each study. Originality/value – The solution combines several technical approaches that are so far separately examined. Each approach is transferred to the innovative domain of industrial inspections and its applicability in this context is verified. New findings give design recommendations for remote workplaces of robot operators.

Most bacterial genomes contain horizontally acquired and transmissible mobile genetic elements, including temperate bacteriophages and integrative and conjugative elements. Little is known about how these elements interact and co-evolved as parts of their host genomes. In many cases, it is not known what advantages, if any, these elements provide to their bacterial hosts. Most strains of Bacillus subtilis contain the temperate phage SPß and the integrative and conjugative element ICEBs1. Here we show that the presence of ICEBs1 in cells protects populations of B. subtilis from predation by SPß, likely providing selective pressure for the maintenance of ICEBs1 in B. subtilis. A single gene in ICEBs1 (yddK, now called spbK for SPß killing) was both necessary and sufficient for this protection. spbK inhibited production of SPß, during both activation of a lysogen and following de novo infection. We found that expression spbK, together with the SPß gene yonE constitutes an abortive infection system that leads to cell death. spbK encodes a TIR (Toll-interleukin-1 receptor)-domain protein with similarity to some plant antiviral proteins and animal innate immune signaling proteins. We postulate that many uncharacterized cargo genes in ICEs may confer selective advantage to cells by protecting against other mobile elements.

Monitoring Xylella fastidiosa is critical for eradicating or at least containing this harmful pathogen. New low-cost and rapid methods for early detection capability are very much needed. Metabolomics may play a key role in diagnosis; in fact, mobile metabolites could avoid errors in sampling due to erratically distributed pathogens. Of the various different mobile signals, we studied dicarboxylic azelaic acid (AzA) which is a key molecule for biotic stress plant response but has not yet been associated with pathogens in olive trees. We found that infected Olea europaea L. plants of cultivars Cellina di Nardò (susceptible to X. fastidiosa) and Leccino (resistant to the pathogen) showed an increase in AzA accumulation in leaf petioles and in sprigs by approximately seven- and sixfold, respectively, compared with plants negative to X. fastidiosa or affected by other pathogens. No statistically significant variation was found between the X. fastidiosa population level and the amount of AzA in either of the plant tissues, suggesting that AzA accumulation was almost independent of the amount of pathogen in the sample. Furthermore, the association of AzA with X. fastidiosa seemed to be reliable for samples judged as potentially false-negative by quantitative polymerase chain reaction (cycle threshold [Ct] > 33), considering both the absolute value of AzA concentration and the values normalized on negative samples, which diverged significantly from control plants. The accumulation of AzA in infected plants was partially supported by the differential expression of two genes (named OeLTP1 and OeLTP2) encoding lipid transport proteins (LTPs), which shared a specific domain with the LTPs involved in AzA activity in systemic acquired resistance in other plant species. The expression level of OeLTP1 and OeLTP2 in petiole samples showed significant upregulation in samples positive to X. fastidiosa of both cultivars, with higher expression levels in positive samples of Cellina di Nardò compared with Leccino, whereas the two transcripts had a low expression level (Ct > 40) in negative samples of the susceptible cultivar. Although the results derived from the quantification of AzA cannot confirm the presence of the erratically distributed X. fastidiosa, which can be definitively assessed by traditional methods, we believe they represent a fast and cheap screening method for large-scale monitoring.

ABSTRACT We have characterized the Pseudomonas putida KT2440 insertion element ISPpu10. This insertion sequence encodes a transposase which exhibits homology to the transposases and specific recombinases of the Piv/Moov family, and no inverted repeats are present at the borders of its left and right ends, thus constituting a new member of the atypical IS110/IS492 family. ISPpu10 was found in at least seven identical loci in the KT2440 genome, and variants were identified having an extra insertion at distinct loci. ISPpu10 always appeared within the core of specific repetitive extragenic palindromic (REP) sequences TCGCGGGTAAACCCG CT CCTAC, exhibiting high target stringency. One intragenic target was found associated with the truncation of a GGDEF/EAL domain protein. After active in vitro transposition to a plasmid-borne target, a duplication of the CT (underlined above) at the junction as a consequence of the ISPpu10 insertion was experimentally demonstrated for the first time in the IS110/IS492 family. The same duplication was observed after transposition of ISPpu10 from a plasmid to the chromosome of P. putida DOT-T1E, an ISPpu10-free strain with REPs similar to those of strain KT2440. Plasmid ISPpu10-mediated rearrangements were observed in vivo under laboratory conditions and in the plant rhizosphere.

Implicit (‘unconscious’) approach–avoidance tendencies towards stimuli can be measured using the Approach Avoidance Task (AAT). We recently expanded a toolbox for analyzing the raw data of a novel, mobile version of the AAT (mAAT), that asks participants to move their phone towards their face (pull) or away (push) in response to images presented on the phone. We here tested the mAAT reaction time and the mAAT distance in a study with 71 Dutch participants that were recruited online and performed an experiment without coming to the laboratory. The participants used both the mAAT and (explicit) rating scales to respond to photographic images of food. As hypothesized, the rated wanting, rated valence and mAAT reaction time indicated a preference for palatable over unpalatable food, and for Dutch over Asian food. Additionally, as expected, arousal was rated higher for unpalatable than for palatable food, and higher for Dutch than for Asian food. The mAAT distance indicated that the unpalatable food images were moved across larger distances, regardless of the movement direction (pull or push), compared to the palatable food images; and the Dutch food images were moved across larger distances than the Asian food images. We conclude that the mAAT can be used to implicitly probe approach–avoidance motivation for complex images in the food domain. The new measure of mAAT distance may be used as an implicit measure of arousal. The ratings and the mAAT measures do not reflect the exact same information and may complement each other. Implicit measures, such as mAAT variables, are particularly valuable when response biases that can occur when using explicit ratings are expected.

Soil volumetric water content (θ) is a parameter describing one of the most important factors conditioning proper plant growth. Monitoring soil moisture is of particular importance in the rational use of water resources for irrigation, especially during periods of water scarcity. This paper presents a method of measuring soil moisture in the vicinity of the plant root system by means of a probe designed to be mounted on a mobile device used for precise plant irrigation. Due to the specific field conditions of the measurement, the design of the probe was proposed as a monopole antenna. Electromagnetic simulations of the probe were carried out with Ansys HFSS software to optimise its dimensions. Then a prototype of the probe was manufactured to conduct laboratory measurements with the use of a vector network analyser (VNA) working in the 20 kHz to 8 GHz frequency range. The VNA analyser was configured to work in the time-domain reflectometry (TDR) mode. From measurements of the time distance between reflections from the probe’s elements it is possible to calculate the bulk dielectric permittivity of the soil surrounding the probe. Next, based on commonly used soil moisture dielectric calibrations one can determine θ of the soil sample. The paper presents simulation results and laboratory tests of an antenna probe. Due to its tough and durable design, this type of probe gives the possibility of easy application in field conditions, which makes it especially suitable for mechanically demanding measurement systems. As the sensitivity zone is comparatively large, this probe is well-suited to measuring soil moisture in the vicinity of the plant root system.

ABSTRACT Three insertion elements were previously found in a family of germ line-limited mobile elements, the Tlr elements, in the ciliate Tetrahymena. Each of the insertions contains an open reading frame (ORF). Sequence analysis of the deduced proteins encoded by the elements suggests that they are homing endonucleases. The genes are designated TIE1-1, TIE2-1, and TIE3-1 for Tetrahymena insertion-homing endonuclease. The endonuclease motif occupies the amino terminal half of each TIE protein. The C-terminal regions of the proteins are similar to the APETELA2 DNA binding domain of plant transcription factors. The TIE1 and TIE3 elements belong to families of repeated sequences in the germ line micronuclear genome. Comparison of the genes and the deduced proteins they encode suggests that there are at least two distinct families of homing endonuclease genes, each of which appears to be preferentially associated with a specific region of the Tlr elements. The TIE1 and TIE3 elements and their cognates undergo programmed elimination from the developing somatic macronucleus of Tetrahymena. The possible role of homing endonuclease-like genes in the DNA breakage step in developmentally programmed DNA elimination in Tetrahymena is discussed.

The article presents the results of a three-year (2016–2018) study on the role of seeding rates taking into account the depth of seeding on different backgrounds of mineral nutrition in the conditions of gray forest soils in the middle Volga region. Experiments were conducted on the pilot field of the Department of Plant Industry and Horticulture of Kazan State Automobile University. Soil of the experimental site is characterized by the following agrochemical indicators: humus content – 2.9–3.2 % (according to Tyurin), the sum of the absorbed bases – 27 mg.-per 100 g of soil, light hydrolysable nitrogen – 79.0...110 mg/1000 g (according to Corinth), mobile phosphorus and potassium (according to Kirsanov) – 105...184 and 79...149 mg/1000 g of soil, pHsol.– 5.6–5.7. Research material – double-grain wheat Triticum Dicoccum Schrank (spelt) grade Runo. During all the years of research, the soil layer 4 and 6 cm was the most wetted during the sprouting phase. On average, for 3 years the best completeness of sprouting was provided by sowing rates of 4–5 million germplasm seeds per hectare at the depth of seed placement by 2 and 4 cm. Safety of plants to full ripeness at sowing of 6 million grains per hectare on both feeding backgrounds made up at planting of seeds on 2 cm – 414–423 pieces/m2, 4 cm – 411–432 and 6 cm – 356–374 plants on 1 m2. The highest yield for 3 years on average on both feeding grounds (1.93...2.55 t/ha) was provided by sowing 4 million grains per hectare when planting seeds by 4 cm.

The model pathogen Ralstonia solanacearum GMI1000 is the causal agent of the bacterial wilt disease that attacks many solanaceous plants and other hosts but not tobacco (Nicotiana spp.). We found that two type III secretion system effector genes, avrA and popP1, are limiting the host range of strain GMI1000 on at least three tobacco species (N. tabacum, N. benthamiana, and N. glutinosa). Both effectors elicit the hypersensitive response (HR) on these tobacco species, although in different manners; AvrA is the major determinant recognized by N. tabacum and N. benthamiana, while PopP1 appears to be the major HR elicitor on N. glutinosa. Only the double inactivation of the avrA and popP1 genes allowed GMI1000 to wilt tobacco plants, thus showing that GMI1000 intrinsically possesses the functions necessary to wilt tobacco plants. A focused analysis on AvrA revealed that the first 58 N-terminal amino acids are sufficient to direct its injection into plant cells. We identified a hypervariable region in avrA, which contains variable numbers of tandem repeats (VNTR), each composed of 12 base pairs. We show that an 18–amino acid region in which the VNTR insertion occurs is an important domain involved in HR elicitation on N. benthamiana. avrA appears to be the target of various DNA insertions or mobile elements that probably allow R. solanacearum to evade the recognition and defense responses of tobacco.

Fertigation supplies water and liquid fertilizer through the same channel to plants. Using drip irrigation set up for fertigation allows the root zone of the plant to be continuously supplied with nutrients and water throughout the farm season. Conventionally, fertigated systems are controlled using pre-set timers to turn on and off fertilizer injectors and irrigation pumps, and also to set the frequency and duration of supply. Therefore, fertigation management is usually based only on predictive algorithms or historical data, which may not be accurate for all situations. Development of a microcontroller-based fertigation management system within the Nigerian (Sub-Saharan Africa) region using a capacitive soil moisture sensor and a JXCT-IOT Frequency Domain Reflectometry (FDR) soil nitrogen sensor is presented. The sensors are placed in the soil around the root region of plants to enable a microcontroller to monitor the soil properties, determine how much water or nutrients the plant needs, and supply the amount needed through a drip irrigation framework. The tap water and urea solution are placed inside separate reservoirs and supplied to the plant through solenoid valves controlled by the microcontroller. Furthermore, an Internet of Things (IoT) client (Blynk IoT) was integrated with the fertigation system so that the fertigation process, as well as the soil state, could be monitored and controlled remotely. The data read from the sensors as well as the state of the solenoid valves were sent over the internet to be stored on the Blynk servers. A website and mobile (Android) dashboards were also created using the Blynk IOT platform to display the states of the valves and the sensor readings. The automatic fertigation system was found to be functional. The system keeps the soil moisture and nitrogen content between the recommended ranges: moisture content between 25% and 46% and nitrogen content between 20 mg/kg and 30 mg/kg for cucumber crops. Fertigation events occur every morning between 5 and 6 am.

Automated identification of plants and animals has improved considerably in the last few years, in particular thanks to the recent advances in deep learning. In order to evaluate the performance of automated plant identification technologies in a sustainable and repeatable way, a dedicated system-oriented benchmark was setup in 2011 in the context of ImageCLEF (Goëau et al. 2011). Each year, since that time, several research groups participated in this large collaborative evaluation by benchmarking their image-based plant identification systems. In 2014, the LifeCLEF research platform (Joly et al. 2014) was created in the continuity of this effort so as to enlarge the evaluated challenges by considering birds and fishes in addition to plants, and audio and video contents in addition to images. The 2017-th edition of the LifeCLEF plant identification challenge (Joly et al. 2017) is an important milestone towards automated plant identification systems working at the scale of continental floras with 10.000 plant species living mainly in Europe and North America illustrated by a total of 1.1M images. Nowadays, such ambitious systems are enabled thanks to the conjunction of the dazzling recent progress in image classification with deep learning and several outstanding international initiatives, aggregating the visual knowledge on plant species coming from the main national botanical institutes. The PlantCLEF plant challenge that we propose to present at this workshop aimed at evaluating to what extent a large noisy training dataset collected through the web (then containing a lot of labelling errors) can compete with a smaller but trusted training dataset checked by experts. To fairly compare both training strategies, the test dataset was created from a third data source, the Pl@ntNet (Joly et al. 2015) mobile application that collects millions of plant image queries all over the world. Due to the good results obtained at the 2017-th edition of the LifeCLEF plant identification challenge, the next big question is how far such automated systems are from the human expertise. Indeed, even the best experts are sometimes confused and/or disagree with each other when validating images of living organism. A multimedia data actually contains only partial information that is usually not sufficient to determine the right species with certainty. Quantifying this uncertainty and comparing it to the performance of automated systems is of high interest for both computer scientists and expert naturalists. This work reports an experimental study following this idea in the plant domain. In total, 9 deep-learning systems implemented by 3 different research teams were evaluated with regard to 9 expert botanists of the French flora. The main outcome of this work is that the performance of state-of-the-art deep learning models is now close to the most advanced human expertise. This shows that automated plant identification systems are now mature enough for several routine tasks, and can offer very promising tools for autonomous ecological surveillance systems.

Motile bacteria follow gradients of nutrients or other environmental cues. Many bacterial chemoreceptors that sense exogenous amino acids contain a double Cache (dCache; calcium channels and chemotaxis receptors) ligand-binding domain (LBD). A growing number of studies suggest that broad-specificity dCache-type receptors that sense more than one amino acid are common. Here, we present an investigation into the mechanism by which the dCache LBD of the chemoreceptor CtaA from a plant growth–promoting rhizobacterium, Pseudomonas fluorescens, recognizes several chemically distinct amino acids. We established that amino acids that signal by directly binding to the CtaA LBD include ones with aliphatic (l-alanine, l-proline, l-leucine, l-isoleucine, l-valine), small polar (l-serine), and large charged (l-arginine) side chains. We determined the structure of CtaA LBD in complex with different amino acids, revealing that its ability to recognize a range of structurally and chemically distinct amino acids is afforded by its easily accessible plastic pocket, which can expand or contract according to the size of the ligand side chain. The amphipathic character of the pocket enables promiscuous interactions with both polar and nonpolar amino acids. The results not only clarify the means by which various amino acids are recognized by CtaA but also reveal that a conserved mobile lid over the ligand-binding pocket adopts the same conformation in all complexes, consistent with this being an important and invariant part of the signaling mechanism.

Enterococci are important opportunistic pathogens with the capacity to acquire and spread antibiotic resistance. At present, linezolid-resistant enterococci (LRE) pose a great challenge. Linezolid is considered as a last resort antibiotic in the treatment of enterococcal infections, so it is important to monitor the occurrence of LRE in various environments. The aim of this study was to define the genetic mechanisms of linezolid resistance in enterococci (E. faecalis, E. faecium, E. hirae, E. casseliflavus) isolated from foods of animal origin (n = 104). Linezolid resistance (LR) was shown by 26.9% of isolates. All of them displayed linezolid MICs of 8–32 µg/mL, and 96.4% of them were multidrug multidrug-resistant. The most common acquired linezolid resistance gene in LR isolates was poxtA (64%), followed by optrA (28%) and cfr (12%). According to the authors’ knowledge, this research is the first to indicate the presence of the cfr gene among isolates from food. In 28.6% of the isolates, the point mutation G2576T in the V domain of the 23S rRNA was responsible for linezolid resistance. All isolates harbored the wild-type rplC, rplD and rplV genes. The obtained results indicate that linezolid resistance among enterococci in animal-derived food may result from various genetic mechanisms. The most worrying is that this resistance is encoded on mobile genetic elements, so there is a risk of its rapid transmission, even despite the lack of selective pressure resulting from the use of antibiotics.

In India, crop area is largest in the world and produces major crops like wheat, pulses, fruits, rice and vegetables Despite of using modem taming techniques along with traditional, infectious plant diseases is major problem which can be caused by different viruses, fungus and bacteria. This mainly affects crop production as well as crop quality. It is very important to identify diseases at early stage Nowadays, automatic crop de detection has become a important research domain. It helps in detecting the symptoms of the disease when they are found on the e In this paper we will focus on finding the diseases in order to increase crop quality and production effectively. Here, we will focus on r diseases by observing leaves of plants at initial stage using machine learning. · In this paper, we designed a Deep Convolutional Neural Network based on LeNet to perform soybean leaf spot disease recognition and classification using affected areas of disease spots. The affected areas of disease spots were segmented from the leaves images using the Unsupervised fuzzy clustering algorithm. The proposed Deep Convolutional Neural Network model achieved a testing accuracy of 89.84%, and poor per class recognition results in 1378 images misclassified, and 1271 images correct classified. TheVGG16 achieved the best performance reaching a 93.54% success rate, and better per class recognition results in 1245 images misclassified, and 1404 images correct classified · In order to address the challenges related to the classification and recognition of soybean disease and healthy leaf identification, it is essential to have access to high-quality images. A meticulously curated dataset named “SoyNet” has been created to provide a clean and comprehensive dataset for research purposes. The dataset comprises over 9000 highquality soybean images, encompassing healthy and diseased leaves. These images have been captured from various angles and directly sourced from soybean agriculture fields; The soybean leaves images are organized into two sub-folders: SoyNet Raw Data and SoyNet Pre-processing Data.The SoyNet Pre-processing Data folder comprises resized images of 256∗256 pixels and the grayscale versions of disease and healthy images, following a similar organizational structure. We captured the images using the Nikon digital camera and the Motorola mobile phone camera, utilizing different angles, lighting conditions, and backgrounds. They were taken in different lighting conditions and backgrounds at soybean cultivation fields to represent the real-world scenario accurately. The proposed dataset is valuable for testing, training, and validating soybean leaf disease classification

This PhD dissertation deals with how to design mobile technology to support mobile work. More specifically, I have worked with the following three connected components: design (as process as well as tool), mobile work (specifically on process plants), and technology (mobile and stationary and the relationship between them). <br /> I define the relationship between these three components based on the action-oriented research approach known as the Scandinavian tradition for systems development, which has grounded and shaped my research method. Cooperation, active user involvement and experimental development of methods and techniques are key elements in this approach, and are elements I develop further in a mobile work environment. Another influencing factor on my approach to the mobile work domain is the current technical development: of the staggering number of microchips produced annually, only a small fraction is being used for Personal Computers the rest are used in mobile and embedded technology. Thus we are dealing with an entirely new range of technology, possibilities and limitations, and consequently new user interface challenges. <br /> Based on this, I see the following key connections to design of technological support of mobile work in a process setting: the design process and the usability methods we use create an understanding of the work practice and the work situations we are trying to support with new technology. The work practise and specific tasks set the boundaries for what we support and how we go about it, and particularly in a mobile work context it is important to understand the heterogeneity in the work: the work tasks and the available resources change in relation to where you are (at the home office, at the site of a client, amongst the machines in the plant) and the users' needs for technological support and access to information changes accordingly. These relationships must be reflected in the technology we design, and it is thus often in a mobile work context more relevant to create a selection of different technological tools for the users to choose between as their needs change, than by replacing one technological approach or device with another. This bears particular importance in relation to mobile technology as the handheld device often supports the mobility but sacrifices the sense of overview because the physically small screens are unable to support it as well as a large PC monitor. <br /> Consequently, the relationship between the three elements is an important aspect of my PhD dissertation. I deal with how mobile work makes new demands on the technological devices we design to support it, thereof mobile technology. I also deal with how the mobile artefacts demand new thinking in relation to the user interface design and functionality because we are dealing with a new type of technology, not just a very small PC with limited screen and next to no peripherals. Finally, I deal with how this new area of research (and the development of HCI and usability in general) sets new demands for the usability methods, how and to what degree we involve users and other professionals in design, how we take advantage of the use environment and utilise the use context actively in the design process. <br /> As a collective concept for the development of mobile technology to support mobile work, I introduce the `web-of-technology' concept that requires we create an understanding of which role the new device should play in relation to the already existing technology in the specific work domain. By placing the new, hand-held technology in the existing `web-of-technology' you are forced to analyse which relations the mobile device should have to the other technological devices in the work context and whether the mobile artefact should be strongly, weakly, or not integrated with the other technology at all. The degree of integration between technologies affects the design of both the functionality and the user interface; strongly integrated devices demand a large degree of visual and functional consistency across devices. It is thus essential that we clarify these questions of the technology that is and the technology that will be early in the design process, just as naturally as we examine how the work practice affects the development and design.

Abstract Rho family proteins are central to the regulation of cell polarity in eukaryotes. Rho of Plants-Guanyl nucleotide Exchange Factor (ROPGEF) can form self-organizing polar domains following co-expression with an Rho of Plants (ROP) and an ROP GTPase-Activating Protein (ROPGAP). Localization of ROPs in these domains has not been demonstrated, and the mechanisms underlying domain formation and function are not well understood. Here we show that six different ROPs form self-organizing domains when co-expressed with ROPGEF3 and GAP1 in Nicotiana benthamiana or Arabidopsis (Arabidopsis thaliana). Domain formation was associated with ROP–ROPGEF3 association, reduced ROP mobility, as revealed by time-lapse imaging and Fluorescence Recovery After Photobleaching beam size analysis, and was independent of Rho GTP Dissociation Inhibitor mediated recycling. The domain formation depended on the ROPs’ activation/inactivation cycles and interaction with anionic lipids via a C-terminal polybasic domain. Coexpression with the microtubule-associated protein ROP effector INTERACTOR OF CONSTITUTIVELY ACTIVE ROP 1 (ICR1) revealed differential function of the ROP domains in the ability to recruit ICR1. Taken together, the results reveal mechanisms underlying self-organizing ROP domain formation and function.

Abstract Phased secondary small interfering RNAs (phasiRNAs) in plants play important roles in regulating genome stability, plant development and stress adaption. Camellia sinensis var. assamica has immense economic, medicinal and cultural significance. However, there are still no studies of phasiRNAs and their putative functions in this valuable plant. We identified 476 and 43 PHAS loci which generated 4290 twenty one nucleotide (nt) and 264 twenty four nt phasiRNAs, respectively. Moreover, the analysis of degradome revealed more than 35000 potential targets for these phasiRNAs. We identified several conserved 21 nt phasiRNA generation pathways in tea plant, including miR390 → TAS3, miR482/miR2118 → NB-LRR, miR393 → F-box, miR828 → MYB/TAS4, and miR7122 → PPR in this study. Furthermore, we found that some transposase and plant mobile domain genes could generate phasiRNAs. Our results show that phasiRNAs target genes in the same family in cis- or trans-manners, and different members of the same gene family may generate the same phasiRNAs. The phasiRNAs, generated by transposase and plant mobile domain genes, and their targets, suggest that phasiRNAs may be involved in the inhibition of transposable elements in tea plant. To summarize, these results provide a comprehensive view of phasiRNAs in Camellia sinensis var. assamica.

This research explores the intersection of Java programming and plant genetic resources (PGR) management by focusing on the development and implementation of the NBPGR-PDS (National Bureau of Plant Genetic Resources Passport Data Sheet) app. By utilizing Java in mobile development, the NBPGR-PDS app addresses challenges in real-time data collection, security, and usability during field explorations for plant germplasm. This paper investigates how Java’s core features—such as platform independence, multithreading, and security—support scientific research in PGR, ensuring reliable data management and enhancing germplasm collection missions. The study concludes with recommendations for improving digital tools and future research directions in this domain.

Abstract This article explores the optimization of plant characteristics and controller parameters for electrified mobility. Electrification of mobile transportation systems, such as automobiles and aircraft, presents the ability to improve key performance metrics such as efficiency and cost. However, the strong bidirectional coupling between electrical and thermal dynamics within new components creates integration challenges, increasing component degradation, and reducing performance. Diminishing these issues requires novel plant designs and control strategies. The electrified mobility literature provides prior studies on plant and controller optimization, known as control co-design (CCD). A void within these studies is the lack of model predictive control (MPC), recognized to manage multi-domain dynamics for electrified systems, within CCD frameworks. This article addresses this through three contributions. First, a thermo-electromechanical hybrid electric vehicle (HEV) powertrain model is developed that is suitable for both plant optimization and MPC. Second, simultaneous plant and controller optimization is performed for this multi-domain system. Third, MPC is integrated within a CCD framework using the candidate HEV powertrain model. Results indicate that optimizing both the plant and MPC parameters simultaneously can reduce physical component sizes by over 60% and key performance metric errors by over 50%.

In current era, Deep Convolution Neural Networks (DCNNs) are desperately improved localization, identification and detection of objects. Recent days, Big data is evolved which leads huge data generation through modern tools like surveillance video cameras. In this paper, we have focused on plant data images in agricultural field. Agriculture is one of major living source in India. To increase the yield by preventing diseases and detection of diseases place major role in agriculture domain. By using Improved and customized DCNN model (improved-detect), We trained plantdoc and plant village datasets. Mainly we used Tomato, Corn and potato plant for model training and testing. we have experimented on plant image data set-tomato leaves both healthy and diseased ones. Experimental results are compared with state of the architectures like Mobile Net, Dark Net-19, ResNet-101and proposed model out PERFORMS in location and detection of plant diseases. obtains best results in computation and accuracy. In the below results sections, we have presented the results with suitable models.

AbstractField measurements of hydrologic tracers indicate varying magnitudes of geochemical separation between subsurface pore waters. The potential for conventional soil physics alone to explain isotopic differences between preferential flow and tightly-bound water remains unclear. Here, we explore physical drivers of isotopic separations using 650 different model configurations of soil, climate, and mobile/immobile soil-water domain characteristics, without confounding fractionation or plant uptake effects. We find simulations with coarser soils and less precipitation led to reduced separation between pore spaces and drainage. Amplified separations are found with larger immobile domains and, to a lesser extent, higher mobile-immobile transfer rates. Nonetheless, isotopic separations remained small (<4‰ for δ2H) across simulations, indicating that contrasting transport dynamics generate limited geochemical differences. Therefore, conventional soil physics alone are unlikely to explain large ecohydrological separations observed elsewhere, and further efforts aimed at reducing methodological artifacts, refining understanding of fractionation processes, and investigating new physiochemical mechanisms are needed.

AbstractAchieving a robust long‐term deployment with mobile robots in the agriculture domain is both a demanded and challenging task. The possibility to have autonomous platforms in the field performing repetitive tasks, such as monitoring or harvesting crops, collides with the difficulties posed by the always‐changing appearance of the environment due to seasonality. With this scope in mind, we report an ongoing effort in the long‐term deployment of an autonomous mobile robot in a vineyard, with the main objective of acquiring what we called the Bacchus Long‐Term (BLT) data set. This data set consists of multiple sessions recorded in the same area of a vineyard but at different points in time, covering a total of 7 months to capture the whole canopy growth from March until September. The multimodal data set recorded is acquired with the main focus put on pushing the development and evaluations of different mapping and localization algorithms for long‐term autonomous robots operation in the agricultural domain. Hence, besides the data set, we also present an initial study in long‐term localization using four different sessions belonging to four different months with different plant stages. We identify that state‐of‐the‐art localization methods can only cope partially with the amount of change in the environment, making the proposed data set suitable to establish a benchmark on which the robotics community can test its methods. On our side, we anticipate two solutions pointed at extracting stable temporal features for improving long‐term 4D localization results. The BLT data set is available at https://lncn.ac/lcas-blt.

Herbal medicinal plant is a traditional medicinal plant that is used to cure a disease. Most of modern people did not know yet the benefits that will be gotten from herbal plants for the health. This research developed a supporting decision application system of herbal medicinal plants for various diseases. ROC (Rank Order Centroid) method was used to count the total number of criteria value and Oreste method was used to rank the alternative herbal medicinal plants with criteria which influence it, namely disease, blood pressure, tall, weight, user’s condition (other diseases), age, kinds of plants, substance and efficacy of plants themselves. Final result of this system was that there were some alternative herbal medicinal plants which were appropriate to user’s disease. In this research, the researcher conducted white box testing by using path base testing to make complex logical estimates to define current action and conducted black box testing by using equivalence partitioning technique which divided domain input, decided testing case by explaining kinds of mistakes. The results of proper test for the system which were done by using questionnaire were gotten 86.75% for testing of functional system, 87% for interface and accessing testing, and 87.33% for testing of advantages system.

AbstractThe plant nuclear genome is largely composed of mobile DNA, which can rearrange genomes and other individual gene structure and also affect gene regulation through various promoted activities: transposition, insertion, excision, chromosome breakage, and ectopic recombination. Ty1-copia-like retrotransposon is a widespread class of transposable elements in the plant kingdom, representing a large part of the total DNA content. Here, a novel retrotransposon-like sequence was isolated and identified as the Ty1-copia-like reverse transcriptase domain (named here CLCoy1), based on the homology of known elements. Fluorescence in situ hybridization, revealed that CLCoy1 was mainly located in telomeric and sub-telomeric regions along the Citrus chromosomes. CLCoy1 composes 3.6% of the genome and, interestingly, while transposons are mostly specific to a species, this element was identified in other Citrus species such as Citrus aurantium, Fortunella margarita and Citrus paradisi, but undetected in Poncirus trifoliata. We also determined that wounding, salt and cell culture stress produced transcriptional activation of this novel retroelement in Citrus limon. The novel Ty1-copia-like element CLCoy1 may have played a major role in shaping genome structure and size during Citrus species evolution.

The impact of climate change has been alarming for the crop growth. The extreme weather conditions can stress the crops and reduce the yield of major crops belonging to Poaceae family too, that sustains 50% of the world’s food calorie and 20% of protein intake. Computational approaches, such as artificial intelligence-based techniques have become the forefront of prediction-based data interpretation and plant stress responses. In this study, we proposed a novel activation function, namely, Gaussian Error Linear Unit with Sigmoid (SIELU) which was implemented in the development of a Deep Learning (DL) model along with other hyper parameters for classification of unknown abiotic stress protein sequences from crops of Poaceae family. To develop this models, data pertaining to four different abiotic stress (namely, cold, drought, heat and salinity) responsive proteins of the crops belonging to poaceae family were retrieved from public domain. It was observed that efficiency of the DL models with our proposed novel SIELU activation function outperformed the models as compared to GeLU activation function, SVM and RF with 95.11%, 80.78%, 94.97%, and 81.69% accuracy for cold, drought, heat and salinity, respectively. Also, a web-based tool, named DeepAProt (http://login1.cabgrid.res.in:5500/) was developed using flask API, along with its mobile app. This server/App will provide researchers a convenient tool, which is rapid and economical in identification of proteins for abiotic stress management in crops Poaceae family, in endeavour of higher production for food security and combating hunger, ensuring UN SDG goal 2.0.

Advancements in phenotyping technology have enabled plant science researchers to gather large volumes of information from their experiments, especially those that evaluate multiple genotypes. To fully leverage these complex and often heterogeneous data sets (i.e. those that differ in format and structure), scientists must invest considerable time in data processing, and data management has emerged as a considerable barrier for downstream application. Here, we propose a pipeline to enhance data collection, processing, and management from plant science studies comprising of two newly developed open-source programs. The first, called AgTC, is a series of programming functions that generates comma-separated values file templates to collect data in a standard format using either a lab-based computer or a mobile device. The second series of functions, AgETL, executes steps for an Extract-Transform-Load (ETL) data integration process where data are extracted from heterogeneously formatted files, transformed to meet standard criteria, and loaded into a database. There, data are stored and can be accessed for data analysis-related processes, including dynamic data visualization through web-based tools. Both AgTC and AgETL are flexible for application across plant science experiments without programming knowledge on the part of the domain scientist, and their functions are executed on Jupyter Notebook, a browser-based interactive development environment. Additionally, all parameters are easily customized from central configuration files written in the human-readable YAML format. Using three experiments from research laboratories in university and non-government organization (NGO) settings as test cases, we demonstrate the utility of AgTC and AgETL to streamline critical steps from data collection to analysis in the plant sciences.

Using naturally occurring bioactive compounds has been crucial in progressing contemporary medical practices. The rhizosphere, which refers to the narrow region of soil surrounding plant roots, serves as a highly dynamic environment for soil bacteria owing to the substantial quantity of organic compounds released into the soil by plant roots. This paper outlines the fermentation and subsequent processing of rhizosphere Sphingomonas egypticus DM. The culture media of Sphingomonas was cultivated and subsequently subjected to propagation. The ethyl acetate extract was then obtained and subjected to fractionation and purification utilizing various chromatographic techniques. This process led to the isolation of a compound under investigation that exhibited a distinct spot on thin-layer chromatography (TLC) plates, with a calculated retention factor (Rf = 0.56) using a mobile phase composed of methylene chloride and methanol (9:1). Upon spraying with anisaldehyde/sulphuric reagent and subsequent heating for a brief period, the compound displayed a violet colour. This compound was investigated in vitro to assess its antimicrobial and minimum inhibitory concentration (MIC) capabilities. The results demonstrated a significant inhibitory effect against phytopathogenic fungi, specifically Rhizoctonia solani (21.3 mm) and Alternaria alternate (18.3 mm). Additionally, a moderate inhibitory effect was observed against Pseudomonas aeruginosa (12.7 mm), whereas a somewhat less inhibitory effect was shown against Streptococcus mutans (9.3 mm). The study also demonstrated that (MIC) against Fusarium oxysporum was seen at a concentration of 125 µg/mL compared to various conventional antibiotics. Finally, the strain underwent PCR screening to detect PKS and lipopeptide 4'-phosphopantetheinyl transferase sfp genes. The PCR amplification assay demonstrated the presence of genes encoding the KS domain and Surfactin. Furthermore, the sequences of Sphingomonas egypticus DM have been officially recorded in the NCBI GenBank database and can be accessed using the accession codes OR469907 and OR499756.

Given the definition of an innovation as an implementation of a new or significantly improved product (good or service) (Gault, 2018), it is thesis of this paper that many innovations fail to create social impacts due to lack of involvement of targeted beneficiaries. Using an innovation called ITIKI (Masinde, 2014), we demonstrate that since design thinking (DT) emphasizes on empathy, invention, and iteration, its adoption in creating social innovations helps in reversing this trend (Masinde, 2020). One way of applying DT in designing winning products/services is by pursuing four questions focusing on the what-is, what-if, what-wows and what-works (Liedtka and Ogilvie, 2011). At the core of ITIKI is indigenous knowledge, which is defined as accumulation of knowledge passing from generation to generation, and guides communities in almost every aspect of their interactions with their environment (Mafongoya, 2017). IK has been at the heart of the coping mechanisms employed by Africa’s small-scale farmers to adapt to climatic variabilities.Though known to be very accurate in predicting, especially the short-term weather patterns, IK is highly endangered and many of its aspects are currently at the verge of extinction. Therefore, innovative ways of conserving and enhancing IK for the benefit of the small-scale farmers are required. ITIKI is one such solution - ITIKI stands for Information Technology and Indigenous Knowledge with Intelligence (https://itiki.co.za/). ITIKI is an integrated decision support system that combines scientific weather data with IK to predict droughts. It takes the form of an early warning system with three components for: (1) drought knowledge capturing, (2) prediction and monitoring, (3) communication and dissemination of the forecasts. ITIKI is implemented using artificial intelligent algorithms, weather sensors and a mobile application. The disseminated forecasts enable the small-scale farmers make better and informed decisions, such as on when, what, how, and where to plant. The forecasts are sent via the ITIKI mobile application or SMS message. The forecasts are also accessible through a web portal, emails and audio files. The main characteristic of ITIKI that informed the adoption of DT is the use of IK whose ownership, appreciation, understanding and application is solely in the hands of the local indigenous people. This makes it a purely human-cantered problem and for which three DT tools were applied in understanding and modelling IK: (1) brainstorming, (2) storytelling and (3) Prototyping. Besides, the problem that ITIKI addresses fits in the domain of problems favoured by DT because it deals with the hardly understood drought prediction domain and the level of uncertainty in drought prediction is very high. The end result of the above is high rate of adoption of ITIKI in Kenya, Mozambique and South Africa by more than 12,000 individual farmers and up to 11% increase in crop yields. This success is associated with the ability of to derive the unique needs of the target users (Africa’s small-scale farmers) through the systematic steps supported by design thinking.

Home is where one starts from. As we grow older The world becomes stranger, the pattern more complicated Of dead and living. Not the intense moment Isolated, with no before and after, But a lifetime burning in every moment… (Eliot 204) Questions of just what home might mean emerged with unfortunate biting salience during the writing of this article with the vicious attack of a student knocked to the ground by the force of a broken bottle and then kicked mercilessly in the head. If not for the ministrations of a bystander, there would have been one less person on the planet. Such disruptive and distressing incidents shake up our world – not only for the person who experiences the original event but also for those who find themselves as witnesses. Using the given incident as an exemplar, the following paper explores the concept of home in the context of ruptures and breaks for people who inhabit a blended world of the digital and the physical. To focus investigations, the Winnicottian concept of the holding environment provides a novel way of understanding home as a seamless domain of continuity which, in this instance is the worldspace spans the physico-digital divide. Sitting writing a paper about ‘home’ and the manner in which the virtual and the physical worlds are blending, I glanced up and was shocked. It is very easy to sit within the warmth and comfort of academe, especially if you have a nice toasty office in the midst of winter and to postulate about what home might be. Theories and concepts, heater, pc and comfy chair support feelings of being at home, of feeling like you have a place in the world, that you have an academic home, you have a conceptual home and, …just wait a minute… back shortly… just answering an email… and a virtual home, in which you can interact and exist in wholly other ways. The other day, however, I abandoned writing the earlier paper with the disorienting experience of seeing a student at my door, a person who tumbled in amidst a mass of scrambled sentences, bandaged bleeding hands, and a bruised head-kicked face. An overseas student who should have been knocking on my door to tell me that ‘Hey, I’ve finished my exams’ instead arrived to ask for my advice: ‘Someone attacked me the other night and I don’t know what to do.’ Home, at least the home about which I wrote before the shock of meeting a traumatised student, was a concept and reality that had transformed markedly over the last quarter of the twentieth century. It was a concept that in its shifts revealed a parallel between the setting up of share housing and the emergence of virtual/physical world blending. Home, as I construed it was about the move, by people aged up to thirties, who were frequently moving from family homes towards blended environments in which share housing became specific non-related familial space (McNamara & Connell), a space/place replicated by social networking in the domain of the digital. There it was. Leaning on the work of theorists such as Miriam Meyerhoff in relation to communities of practice in a linguistic sense, to the earlier work of Lesley Milroy in relation to social networks, I was set to make an argument that the textual world of the internet and other digital domains was developing in a manner that replicated linguistic – specifically spoken – communities of practice based on speech patterns. Buoyed by the recent discovery of the more recent writing of Line Dubé, Anne Bourhis and Réal Jacob in relation to virtual communities of practice, I was certain that my propositions regarding textual practices had something to offer to the current edition of this journal. Further, my argument would proceed in such a way as to infer that the textual base played out in digital media was advancing into the domain of speech in the physical world to the extent that it was possible to determine who had an active digital life – especially in relation to domains on the net – merely by their vocabulary and their sentence construction. My proposition was that the digital domain had not only blended with the virtual in the manner that Dubé, Bourhis and Jacob suggested, but that textual communication was now a home base for the development of the English language for a broad section of the general populace in English speaking countries. The sudden jar of a physical world shock shook loose the comfortable home of text and theory and challenged what I wrote. What was home for the young student who stood before me? We had spoken of ‘home’ before, of making home in a new country, of how your housemates become your family to a certain extent, of how internet and mobile phones made it easier, how home was really with you wherever you went BUT, with the disaster that was an assault, some of that rhetoric resonated as hollow – rhetoric without substance, cold comfort, no comfort. In this situation, home is a concept tested. Perhaps only in such a context can the boundaries and meanings of home come to the fore. It is to that issue that I will address this version of the paper and for that purpose, I will advance the argument that although there may well be a modified version of home developing for a specific generation or cohort of people, that there remains a need for anchoring in the various domains of engagement. To that end, I will use the theory of psychoanalytic theorist D.W. Winnicott who constructed the concept of the holding environment (Winnicott ‘From Dependence;’ and Seinfeld). This article therefore takes its new springing point from hereon in and starts with a brief exploration of the holding environment by its originating author, reconstructs this as a contextually relevant concept, and then talks into some of the original propositions using the given incident for illustrative purposes. The holding environment as construed by D.W. Winnicott is, under optimal conditions, the first environment that an infant experiences, the warm and caring one provided by a primary caregiver who, for this article will be known as the m/other (“The Concept of the Healthy Individual” 27-28). Within this environment of literal and metaphoric holding, the infant knows nothing other than an all-encompassing domain which includes physical and psychological care, the anticipation and provision of needs, and a titrated introduction to the world of things and people (“From Dependence” 86). From the perspective of the infant and within this circle of holding, the world belongs to the infant and is composed largely of the m/other. Only when there is a break in the continuity of care does the infant notice/perceive a world that is anything other than seamless with her/his own existence. In Winnicott’s schema, if a holding environment operates in an optimal manner, it largely remains invisible (Winnicott, “From Dependence” 86; Winnicott, “The Theory of the Parent-Infant Relationship” 52; Ogden 200). This manner of experiencing the world changes with the developing person so that in adulthood, we experience a range of environments that attend to our various needs, if we are fortunate enough. For example, your office supports your work to a greater or lesser extent and perhaps your partner supports you in a psychological sense, and your personal trainer supports your physical training needs. Other instances of support and holding could include the glasses that support your sight and the car that supports your proclivity for drives in the country and a particular lifestyle. There are therefore, many things, people, institutions, and even phenomena such as birthday celebrations that support different aspects of who we are – our being – and different aspects of our activities – our doing. This mirrors theories developed within the context of sociolinguistics in which authors parallel what people are with social networks and what people do, with communities of practice (Moore 22). In the context of Winnicott and linguistic theory, without those supports, our lives would be different and for many of us, would be diminished. The supports I describe are those I construe as holding environments and I believe that by considering a holding environment as a form of ‘home’ that we can reveal a specific way of understanding not only what a home might be, but also the manner in which it operates when people perceive it to be under threat. In the context of the digital domain, there are many media such as email, chat rooms, twitter, real time chat in a range of venues and digital social networks and virtual worlds that support different aspects of our identities, of things that we want to do, of contacts we make and maintain, and of communication for fun and for business. My initial proposition included the concept that various language forms operate to support and construct our identities and that what digital media provided were various venues for the operation of differing but overlapping holding environments in a textual sense. What do these elements, or those like them mean in the situation in which the student found himself? What does it mean and why was it that despite some time in between, that his primary quest was to seek out a person in the physical domain rather than finding solace online when, as I understood, he spent a great deal of time in digital communication? I believe that although there is a blending of domains – the digital and analogue – that when a holding environment of either variety breaks, fractures or at least reveals cracks, that it is likely that a person will seek redress in both modes and in so doing, will reaffirm what is a vital element for the healthy existence of every person – the maintenance of a sense of home – be that on or offline. Despite the seeking for redress in the mode in which the break occurred, the parallel search for social sanction and acknowledgement in the alternative domain may be just as significant for a slightly different reason. When Winnicott writes about ruptures and breaks, it is about those impingements that destroy continuity (“The Fear of Breakdown” 93) – the break in going on being. In the current context in which a person or community inhabits both the online and offline realms, part of their continuity of being, their worldspace (Hardey 2) is the seamlessness between the domains. It is therefore necessary to bring the sense of rupture/failure that occurs in one domain, across into the other to maintain the meta- holding environment or home. Home is that space where ‘you speak my language,’ whether on or offline, the holding environment is one that adapts to you, that understands your speech/text and responds in a manner predictable and in your own genre under optimal conditions, home meets you where you are and, importantly, is a space and place that when it ruptures, mends in such a way as to your restore your faith in its capacity to perform as a holding environment (“Transitional Objects” 10-11). Winnicott writes that only with an environment that was not perfect, (only with an environment that failed occasionally in a minor way), is it possible for a person to sense that there was a holding environment at all. Further, rather than a person construing this failing as a marker of lack of dependability, that the small failure revealed the significance and value of its effective functioning for most of the time. Additionally, a minor break revealed that the holding environment/home held the potential to respond to some unanticipated and distressing break by supporting the person experiencing it. By operating in this manner, there is now an imaginal space of holding/home. In a sense, this mirrors what other authors such as Thomas Lindif and Milton Shatzer write about when they describe social presence in relation to the manner in which an online arena supports or is perceived to support activities such as communication between peers. One of the most noted and public manifestations of the phenomenon of a failed holding environment becoming mended and therefore stronger was that experienced in several places in relation to terrorist attacks such as that of 2001 in the USA. In relation to the attacks on the twin towers in New York, the people of that city experienced a shattering of the integrity of their holding environment/ their home. However, they also noted – as reported across a range of media (for example: Gamble 1.iii; Grider), a huge outpouring of compassion and caring by their fellow New Yorkers thereby experiencing a certain mending and elevating of the significance of their home city holding environment (Gamble 2.vi). In the context of the aforementioned student being attacked, the break also occurred in the physical domain. Although he sought some form of reassurance online could provide some solace. However, it would leave him with the experience that the physical environment was no longer homelike, that it had failed as a holding environment. That is, home in the physical realm was, for a time, failing to support him. To effect a mending in the physical domain, it was therefore important that he seek out solutions that equally involved the physical world of people – mirroring the break – the assault by a person. What occurred when he visited my office was that he received a physical world hearing and witness to his injuries and then with the aid of colleagues, he received further care, advice and support. One of the consequences of such an experience is that although the possibility of assault is now imaginable, because it has been experienced; there is also the knowledge that assistance is at hand – a situation that may not have been known or predicted before. In some manner therefore, with other imagined ghastly events, there is now an expectation of potential assistance. That imaginal knowing therefore now forms part of his holding environment in his physical world, that form of home that ensures ontological security as mentioned by McNamara and Connell (82). Outrage over incidents in Second Life and in other domains such as myspace predominantly play out in those arenas but, like the assault of the student, also get played out in other arenas, including mainstream media. For example, an attack on the virtual headquarters of the Australian Broadcasting Corporation on Second Life attracted attention in newspapers and other mainstream media (Hutcheon). It seems therefore that not only is it necessary to mend the breaks in a sense within the medium in which the original break occurred but also to reassert the blended domain of the digital and the analogue and the capacity of each to form part of the meta holding environment that exists in contemporary society. There is yet to develop a discourse that links the digital and the physical worlds as constituents of a worldspace (Hardey 2), that can be viewed as a meta- holding environment/home. However, even with the few examples proffered here, it seems apparent that by investigating breaks and ruptures in the lives of people who maintain a life world that spans the digital/physical divide that it might be possible to understand the apparent merging of the two. Further, it may lead to significant observations about the newly emerging worldspace as a holding environment /home in a novel way with leads for the assisting people across the divides that may otherwise have not been considered. The implications for maintaining the seamlessness and continuity of home/holding environment in the instance of natural or person-effected disasters in either domain is the demand for an appropriate response in both. Although this already occurs, it is in an ad hoc manner without a consideration of the significance of mending ruptures and re-enlivening both domains for a sense of ontological security of the worldspace – that is at its very heart, a sense of home. References Dubé, L., A. Bourhis, and R. Jacob. “Towards a Typology of Virtual Communities of Practice.” Interdisciplinary Journal of Information, Knowledge, and Management 1 (2006): 69-93. Eliot, T. S. “East Coker V.” Collected Poems 1909-26. London: Faber, 1974. 202-204. Gamble, Jennifer M. The Aesthetics of Mourning & the Anaesthetics of Trauma: Transformation through Memorial Space. Ph.D. thesis. The University of Sydney, 2006. Grider, Sylvia. “Spontaneous Shrines: A Modern Response to Tragedy and Disaster (Preliminary Observations Regarding the Spontaneous Shrines Following the Terrorist Attacks of September 11, 2001)”. New Directions in Folklore 5 Oct. 2001: 1-10. 1 Dec. 2002 http://www.temple.edu/isllc/newfolk/shrines.html>. Hardey, Mariann. “Going Live: Converging Mobile Technology and the Sociability of the iGeneration.” M/C Journal 10.1 (2007). 2 July 2007 http://journal.media-culture.org.au/0703/09-hardey.php>. Hutcheon, Stephen. “Vandals ‘Bomb’ ABC Island.” Sydney Morning Herald 22 May 2007. 23 May. 2007 http://www.smh.com.au/news/web/vandals-bomb-abc-island/2007/05/22/1179601400256.html>. Lindlif, Thomas R., and Milton J. Shatzer. “Media Ethnography in Virtual Space: Strategies, Limits, and Possibilities.” Journal of Broadcasting & Electronic Media 42.2 (1998): 170(20). McNamara, Sophie, and John Connell. “Homeward Bound? Searching for Home in Inner Sydney’s Share Houses.” Australian Geographer 38.1 (2007): 71-91. Meyerhoff, Miriam. “Communities of Practice.” Handbook of Language Variation and Change. Eds. J.K. Chambers, Natalie Schilling-Estes and Peter Trudgill. Oxford: Basil Blackwell, 2002. 526-548. Milroy, J., and L. Milroy. “Linguistic Change, Social Network and Speaker Innovation.” Journal of Linguistics 21.2 (1985): 229-284. Moore, Emma. Learning Style and Identity: A Sociolinguistic Analysis of a Bolton High School. Unpublished PhD dissertation. Manchester, UK: University of Manchester (2003). Ogden, Thomas H. The Matrix of the Mind: Object Relations and the Psychoanalytic Dialogue. London: Maresfield Library, 1990. Seinfeld, Jeffrey. “Donald Winnicott and the Holding Relationship.” Interpreting and Holding: The Paternal and Maternal Functions of the Psychotherapist. Northvale, New Jersey & London: Jason Aronson, 1993. 101-121. Winnicott, Donald Woods. “The Concept of a Healthy Individual.” D.W. Winnicott: Home Is Where We Start From: Essays by a Psychoanalyst. Eds. Clare Winnicott, Ray Shepherd, and Madeleine Davis. New York: Penguin, 1975 (A talk given to the Royal Medico-Psychological Association, Psychotherapy and Social Psychiatry Section, 8 March 1967). 21-39. ———. “The Fear of Breakdown.” D. W. Winnicott: Psycho-Analytic Explorations. Eds. Clare Winnicott, Ray Shepherd and Madeleine Davis. Vol. 1. London: Karnac Books, 1989 (paper originally written c. 1963). 87-96. ———. “From Dependence towards Independence in the Development of the Individual.” The Maturational Processes and the Facilitating Environment. London: Karnac Books, 2002 (Paper first presented in 1963). 83-92. ———. “The Theory of the Parent-Infant Relationship.” The Maturational Processes and the Facilitating Environment. London: Karnac Books, 2002 (Paper first presented in 1960). 37-55. ———. “Transitional Objects and Transitional Phenomena.” Playing and Reality. London: Brunner-Routledge, 1971/2001. 1-30. Citation reference for this article MLA Style Gamble, Jennifer M. "Holding Environment as Home: Maintaining a Seamless Blend across the Virtual/Physical Divide." M/C Journal 10.4 (2007). echo date('d M. Y'); ?> <http://journal.media-culture.org.au/0708/11-gamble.php>. APA Style Gamble, J. (Aug. 2007) "Holding Environment as Home: Maintaining a Seamless Blend across the Virtual/Physical Divide," M/C Journal, 10(4). Retrieved echo date('d M. Y'); ?> from <http://journal.media-culture.org.au/0708/11-gamble.php>.

The Mangrove ecosystem offers a range of benefits and opportunities for local and national economic development such as fisheries, shoreline stability, and resource sustainability hence they should be protected and conserved. In Kenya Mangroves were declared government reserve forests by the Proclamation No. 44 of 30th April 1932, and later by Legal Notice No. 174 of 20th May 1964. Under this “Gazette Notification for Mangrove Forests in Kenya” all land between high water and low water marks (ordinary spring tides) are described as mangrove areas. These forests cover about 61,279ha representing approximately 3% of the natural forest cover or less than 1% of the national land area. Mangrove forests are found in tidal estuaries, creeks, and protected bays along the 536 km long Kenyan coastline that extends from the Kenya-Tanzania border in the south to the Kenya-Somalia border in the north; between latitudes 1°40΄S and 4°25΄S and longitudes 41°34΄E and 39°17΄E. Mikoko (Mangrove in Swahili) is the first project specifically developed for mangrove areas in Kenya. Driven by the Kenya Forest Service, the project takes into account the recommendations of the National Mangrove Ecosystem Management Plan 2017-2027. For effective management of mangroves in the country, the Plan is organised around six programmes including; forest conservation and utilization; fisheries development and management; community; tourism development; research and education; and human resource and operations. These programmes prescribe measures for rehabilitation, conservation, and sustainable management of mangrove ecosystems in Kenya. Implementations of this Plan will be led by Kenya Forest Service (KFS); but will also include other key actors such as Kenya Wildlife Service, State Department of Fisheries, Research Institutions, Academia and Community Forest Associations through a specially constituted National Mangrove Advisory Committee. The project is for a 2-year period during which it shall develop an effective system for management of mangrove resources in Kenya. This will be done through the development of management prescriptions, skills improvement and provision of adequate infrastructure as recommended by the mangrove plan. Biodiversity information on mangroves, covering species descriptions and distribution, is essential for understanding the ecology and distribution of this endangered ecosystem and its management. Nowadays, very few is done in that domain. If the mangroves location are well know, it's not the case regarding the species distribution. Citizen science is a way to address this important issue. MIKOKO, a French FSPI funded project, is focused on Kenya to develop species information systems on mangrove species. Mikoko aims to assimilate knowledge through its participatory portal to strengthen a science and technology network of stakeholders such as scientists and parataxonomists in the African region. The portal platform will contain a Citizen Science module, a graphic driven species identification module, a species pages module conforming to Species Profile Model and a spatial module. Subscribing to open data paradigm, all the data on the portal will be covered by Creative Commons license framework. Mobile applications for identifying 60 plant species and contributing to citizen science module will be deployed. The oral presentation will expose the portal features and related apps seeking participation from the environmentalists from the African region.

Unlike some current discourse on automediality, this essay eschews most of the analysis concerning the adoption or modification of avatars to deliberately enhance, extend or distort the self. Rather than the automedial enabling of alternative, virtual selves modified by playful, confronting or disarming avatars we concentrate instead on emerging efforts to present the self in hyper-realist, interactive modes. In doing so we ask, what is the relationship between traumatic forms of automediation and the affective impact on and response of the audience? We argue that, while on the one hand there are promising avenues for valuable individual and social engagements with traumatic forms of automediation, there is an overwhelming predominance of suffering as a theme in such virtual depictions, comingled with uncritically asserted promises of empathy, which are problematic as the technology assumes greater mainstream uptake.As Smith and Watson note, embodiment is always a “translation” where the body is “dematerialized” in virtual representation (“Virtually” 78). Past scholarship has analysed the capacity of immersive realms, such as Second Life or online games, to highlight how users can modify their avatars in often spectacular, non-human forms. Critics of this mode of automediality note that users can adopt virtually any persona they like (racial, religious, gendered and sexual, human, animal or hybrid, and of any age), behaving as “identity tourists” while occupying virtual space or inhabiting online communities (Nakamura). Furthermore, recent work by Jaron Lanier, a key figure from the 1980s period of early Virtual Reality (VR) technology, has also explored so-called “homuncular flexibility” which describes the capacity for humans to seemingly adapt automatically to the control mechanisms of an avatar with multiple legs, other non-human appendages, or for two users to work in tandem to control a single avatar (Won et. al.). But this article is concerned less with these single or multi-player online environments and the associated concerns over modifying interactive identities. We are principally interested in other automedial modes where the “auto” of autobiography is automated via Artificial Intelligences (AIs) to convincingly mimic human discourse as narrated life-histories.We draw from case studies promoted by the 2017 season of ABC television’s flagship science program, Catalyst, which opened with semi-regular host and biological engineer Dr Jordan Nguyen, proclaiming in earnest, almost religious fervour: “I want to do something that has long been a dream. I want to create a copy of a human. An avatar. And it will have a life of its own in virtual reality.” As the camera followed Nguyen’s rapid pacing across real space he extolled: “Virtual reality, virtual human, they push the limits of the imagination and help us explore the impossible […] I want to create a virtual copy of a person. A digital addition to the family, using technology we have now.”The troubling implications of such rhetoric were stark and the next third of the program did little to allay such techno-scientific misgivings. Directed and produced by David Symonds, with Nguyen credited as co-developer and presenter, the episode “Meet the Avatars” immediately introduced scenarios where “volunteers” entered a pop-up inner city virtual lab, to experience VR for the first time. The volunteers were shown on screen subjected to a range of experimental VR environments designed to elicit fear and/or adverse and disorienting responses such as vertigo, while the presenter and researchers from Sydney University constantly smirked and laughed at their participants’ discomfort. We can only wonder what the ethics process was for both the ABC and university researchers involved in these broadcast experiments. There is little doubt that the participant/s experienced discomfort, if not distress, and that was televised to a national audience. Presenter Nguyen was also shown misleading volunteers on their way to the VR lab, when one asked “You’re not going to chuck us out of a virtual plane are you?” to which Nguyen replied “I don't know what we’re going to do yet,” when it was next shown that they immediately underwent pre-programmed VR exposure scenarios, including a fear of falling exercise from atop a city skyscraper.The sweat-inducing and heart rate-racing exposures to virtual plank walks high above a cityscape, or seeing subjects haptically viewing spiders crawl across their outstretched virtual hands, all elicited predictable responses, showcased as carnivalesque entertainment for the viewing audience. As we will see, this kind of trivialising of a virtual environment’s capacity for immersion belies the serious use of the technology in a range of treatments for posttraumatic stress disorder (see Rizzo and Koenig; Rothbaum, Rizzo and Difede).Figure 1: Nguyen and researchers enjoying themselves as their volunteers undergo VR exposure Defining AutomedialityIn their pioneering 2008 work, Automedialität: Subjektkonstitution in Schrift, Bild und neuen Medien, Jörg Dünne and Christian Moser coined the term “automediality” to problematise the production, application and distribution of autobiographic modes across various media and genres—from literary texts to audiovisual media and from traditional expression to inter/transmedia and remediated formats. The concept of automediality was deployed to counter the conventional critical exclusion of analysis of the materiality/technology used for an autobiographical purpose (Gernalzick). Dünne and Moser proffered a concept of automediality that rejects the binary division of (a) self-expression determining the mediated form or (b) (self)subjectivity being solely produced through the mediating technology. Hence, automediality has been traditionally applied to literary constructs such as autobiography and life-writing, but is now expanding into the digital domain and other “paratextual sites” (Maguire).As Nadja Gernalzick suggests, automediality should “encourage and demand not only a systematics and taxonomy of the constitution of the self in respectively genre-specific ways, but particularly also in medium-specific ways” (227). Emma Maguire has offered a succinct working definition that builds on this requirement to signal the automedial universally, noting it operates asa way of studying auto/biographical texts (of a variety of forms) that take into account how the effects of media shape the kinds of selves that can be represented, and which understands the self not as a preexisting subject that might be distilled into story form but as an entity that is brought into being through the processes of mediation.Sidonie Smith and Julia Watson point to automediality as a methodology, and in doing so emphasize how the telling or mediation of a life actually shapes the kind of story that can be told autobiographically. They state “media cannot simply be conceptualized as ‘tools’ for presenting a preexisting, essential self […] Media technologies do not just transparently present the self. They constitute and expand it” (Smith and Watson “Virtually Me” 77).This distinction is vital for understanding how automediality might be applied to self-expression in virtual domains, including the holographic avatar dreams of Nguyen throughout Catalyst. Although addressing this distinction in relation to online websites, following P. David Marshall’s description of “the proliferation of the public self”, Maguire notes:The same integration of digital spaces and platforms into daily life that is prompting the development of new tools in autobiography studies […] has also given rise to the field of persona studies, which addresses the ways in which individuals engage in practices of self-presentation in order to form commoditised identities that circulate in affective communities.For Maguire, these automedial works operate textually “to construct the authorial self or persona”.An extension to this digital, authorial construction is apparent in the exponential uptake of screen mediated prosumer generated content, whether online or theatrical (Miller). According to Gernalzick, unlike fictional drama films, screen autobiographies more directly enable “experiential temporalities”. Based on Mary Anne Doane’s promotion of the “indexicality” of film/screen representations to connote the real, Gernalzick suggests that despite semiotic theories of the index problematising realism as an index as representation, the film medium is still commonly comprehended as the “imprint of time itself”:Film and the spectator of film are said to be in a continuous present. Because the viewer is aware, however, that the images experienced in or even as presence have been made in the past, the temporality of the so-called filmic present is always ambiguous” (230).When expressed as indexical, automedial works, the intrinsic audio-visual capacities of film and video (as media) far surpass the temporal limitations of print and writing (Gernalzick, 228). One extreme example can be found in an emergent trend of “performance crime” murder and torture videos live-streamed or broadcast after the fact using mobile phone cameras and FaceBook (Bender). In essence, the political economy of the automedial ecology is important to understand in the overall context of self expression and the governance of content exhibition, access, distribution and—where relevant—interaction.So what are the implications for automedial works that employ virtual interfaces and how does this evolving medium inform both the expressive autobiographical mode and audiences subjectivities?Case StudyThe Catalyst program described above strove to shed new light on the potential for emerging technology to capture and create virtual avatars from living participants who (self-)generate autobiographical narratives interactively. Once past the initial gee-wiz journalistic evangelism of VR, the episode turned towards host Nguyen’s stated goal—using contemporary technology to create an autonomous virtual human clone. Nguyen laments that if he could create only one such avatar, his primary choice would be that of his grandfather who died when Nguyen was two years old—a desire rendered impossible. The awkward humour of the plank walk scenario sequence soon gives way as the enthusiastic Nguyen is surprised by his family’s discomfort with the idea of digitally recreating his grandfather.Nguyen next visits a Southern California digital media lab to experience the process by which 3D virtual human avatars are created. Inside a domed array of lights and cameras, in less than one second a life-size 3D avatar is recorded via 6,000 LEDs illuminating his face in 20 different combinations, with eight cameras capturing the exposures from multiple angles, all in ultra high definition. Called the Light Stage (Debevec), it is the same technology used to create a life size, virtual holocaust survivor, Pinchas Gutter (Ziv).We see Nguyen encountering a life-size, high-resolution 2D screen version of Gutter’s avatar. Standing before a microphone, Nguyen asks a series of questions about Gutter’s wartime experiences and life in the concentration camps. The responses are naturalistic and authentic, as are the pauses between questions. The high definition 4K screen is photo-realist but much more convincing in-situ (as an artifact of the Catalyst video camera recording, in some close-ups horizontal lines of transmission appear). According to the project’s curator, David Traum, the real Pinchas Gutter was recorded in 3D as a virtual holograph. He spent 25 hours providing 1,600 responses to a broad range of questions that the curator maintained covered “a lot of what people want to say” (Catalyst).Figure 2: The Museum of Jewish Heritage in Manhattan presented an installation of New Dimensions in Testimony, featuring Pinchas Gutter and Eva SchlossIt is here that the intersection between VR and auto/biography hybridise in complex and potentially difficult ways. It is where the concept of automediality may offer insight into this rapidly emerging phenomenon of creating interactive, hyperreal versions of our selves using VR. These hyperreal VR personae can be questioned and respond in real-time, where interrogators interact either as casual conversers or determined interrogators.The impact on visitors is sobering and palpable. As Nguyen relates at the end of his session, “I just want to give him a hug”. The demonstrable capacity for this avatar to engender a high degree of empathy from its automedial testimony is clear, although as we indicate below, it could simply indicate increased levels of emotion.Regardless, an ongoing concern amongst witnesses, scholars and cultural curators of memorials and museums dedicated to preserving the history of mass violence, and its associated trauma, is that once the lived experience and testimony of survivors passes with that generation the impact of the testimony diminishes (Broderick). New media modes of preserving and promulgating such knowledge in perpetuity are certainly worthy of embracing. As Stephen Smith, the executive director of the USC Shoah Foundation suggests, the technology could extendto people who have survived cancer or catastrophic hurricanes […] from the experiences of soldiers with post-traumatic stress disorder or survivors of sexual abuse, to those of presidents or great teachers. Imagine if a slave could have told her story to her grandchildren? (Ziv)Yet questions remain as to the veracity of these recorded personae. The avatars are created according to a specific agenda and the autobiographical content controlled for explicit editorial purposes. It is unclear what and why material has been excluded. If, for example, during the recorded questioning, the virtual holocaust survivor became mute at recollecting a traumatic memory, cried or sobbed uncontrollably—all natural, understandable and authentic responses given the nature of the testimony—should these genuine and spontaneous emotions be included along with various behavioural ticks such as scratching, shifting about in the seat and other naturalistic movements, to engender a more profound realism?The generation of the photorealist, mimetic avatar—remaining as an interactive persona long after the corporeal, authorial being is gone—reinforces Baudrillard’s concept of simulacra, where a clone exists devoid of its original entity and unable to challenge its automedial discourse. And what if some unscrupulous hacker managed to corrupt and subvert Gutter’s AI so that it responded antithetically to its purpose, by denying the holocaust ever happened? The ethical dilemmas of such a paradigm were explored in the dystopian 2013 film, The Congress, where Robyn Wright plays herself (and her avatar), as an out of work actor who sells off the rights to her digital self. A movie studio exploits her screen persona in perpetuity, enabling audiences to “become” and inhabit her avatar in virtual space while she is limited in the real world from undertaking certain actions due to copyright infringement. The inability of Wright to control her mimetic avatar’s discourse or action means the assumed automedial agency of her virtual self as an immortal, interactive being remains ontologically perplexing.Figure 3: Robyn Wright undergoing a full body photogrammetry to create her VR avatar in The Congress (2013)The various virtual exposures/experiences paraded throughout Catalyst’s “Meet the Avatars” paradoxically recorded and broadcast a range of troubling emotional responses to such immersion. Many participant responses suggest great caution and sensitivity be undertaken before plunging headlong into the new gold rush mentality of virtual reality, augmented reality, and AI affordances. Catalyst depicted their program subjects often responding in discomfort and distress, with some visibly overwhelmed by their encounters and left crying. There is some irony that presenter Ngyuen was himself relying on the conventions of 2D linear television journalism throughout, adopting face-to-camera address in (unconscious) automedial style to excitedly promote the assumed socio-cultural boon such automedial VR avatars will generate.Challenging AuthenticityThere are numerous ethical considerations surrounding the potential for AIs to expand beyond automedial (self-)expression towards photorealist avatars interacting outside of their pre-recorded content. When such systems evolve it may be neigh impossible to discern on screen whether the person you are conversing with is authentic or an indistinguishable, virtual doppelganger. In the future, a variant on the Turning Test may be needed to challenge and identify such hyperreal simulacra. We may be witnessing the precursor to such a dilemma playing out in the arena of audio-only podcasts, with some public intellectuals such as Sam Harris already discussing the legal and ethical problems from technology that can create audio from typed text that convincingly replicate the actual voice of a person by sampling approximately 30 minutes of their original speech (Harris). Such audio manipulation technology will soon be available to anybody with the motivation and relatively minor level of technological ability in order to assume an identity and masquerade as automediated dialogue. However, for the moment, the ability to convincingly alter a real-time computer generated video image of a person remains at the level of scientific innovation.Also of significance is the extent to which the audience reactions to such automediated expressions are indeed empathetic or simply part of the broader range of affective responses that also include direct sympathy as well as emotions such as admiration, surprise, pity, disgust and contempt (see Plantinga). There remains much rhetorical hype surrounding VR as the “ultimate empathy machine” (Milk). Yet the current use of the term “empathy” in VR, AI and automedial forms of communication seems to be principally focused on the capacity for the user-viewer to ameliorate negatively perceived emotions and experiences, whether traumatic or phobic.When considering comments about authenticity here, it is important to be aware of the occasional slippage of technological terminology into the mainstream. For example, the psychological literature does emphasise that patients respond strongly to virtual scenarios, events, and details that appear to be “authentic” (Pertaub, Slater, and Barker). Authentic in this instance implies a resemblance to a corresponding scenario/activity in the real world. This is not simply another word for photorealism, but rather it describes for instance the experimental design of one study in which virtual (AI) audience members in a virtual seminar room designed to treat public speaking anxiety were designed to exhibit “random autonomous behaviours in real-time, such as twitches, blinks, and nods, designed to encourage the illusion of life” (Kwon, Powell and Chalmers 980). The virtual humans in this study are regarded as having greater authenticity than an earlier project on social anxiety (North, North, and Coble) which did not have much visual complexity but did incorporate researcher-triggered audio clips of audience members “laughing, making comments, encouraging the speaker to speak louder or more clearly” (Kwon, Powell, and Chalmers 980). The small movements, randomly cued rather than according to a recognisable pattern, are described by the researchers as creating a sense of authenticity in the VR environment as they seem to correspond to the sorts of random minor movements that actual human audiences in a seminar can be expected to make.Nonetheless, nobody should regard an interaction with these AIs, or the avatar of Gutter, as in any way an encounter with a real person. Rather, the characteristics above function to create a disarming effect and enable the real person-viewer to willingly suspend their disbelief and enter into a pseudo-relationship with the AI; not as if it is an actual relationship, but as if it is a simulation of an actual relationship (USC). Lucy Suchman and colleagues invoke these ideas in an analysis of a YouTube video of some apparently humiliating human interactions with the MIT created AI-robot Mertz. Their analysis contends that, while it may appear on first glance that the humans’ mocking exchange with Mertz are mean-spirited, there is clearly a playfulness and willingness to engage with a form of AI that is essentially continuous with “long-standing assumptions about communication as information processing, and in the robot’s performance evidence for the limits to the mechanical reproduction of interaction as we know it through computational processes” (Suchman, Roberts, and Hird).Thus, it will be important for future work in the area of automediated testimony to consider the extent to which audiences are willing to suspend disbelief and treat the recounted traumatic experience with appropriate gravitas. These questions deserve attention, and not the kind of hype displayed by the current iteration of techno-evangelism. Indeed, some of this resurgent hype has come under scrutiny. From the perspective of VR-based tourism, Janna Thompson has recently argued that “it will never be a substitute for encounters with the real thing” (Thompson). Alyssa K. Loh, for instance, also argues that many of the negatively themed virtual experiences—such as those that drop the viewer into a scene of domestic violence or the location of a terrorist bomb attack—function not to put you in the position of the actual victim but in the position of the general category of domestic violence victim, or bomb attack victim, thus “deindividuating trauma” (Loh).Future work in this area should consider actual audience responses and rely upon mixed-methods research approaches to audience analysis. 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