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The Artificial Gravity Bed Rest with European Space Agency (AGBRESA) study involved 2 campaigns of 60-day bed rest at 6° head down tilt in the:envihab facility in Cologne, Germany, in 2019. The objective was to determine whether centrifugation of supine subjects mitigates physiological changes that occur due to the exposure to the spaceflight analog of 6° head down tilt. A set of international standard measures was used to evaluate bone, muscle, nutritional status, ocular changes, and psychological state and to assess cardiovascular, sensorimotor, and immune systems functions. After 60 days of bed rest, the subjects who were not exposed to centrifugation had significant decreases in mineral density at the hip and lumbar spine, decreased muscle strength of the knee and ankle, reduced maximal aerobic capacity, orthostatic intolerance, and impaired balance and locomotion, as well as significant biochemical, immunological, and psychological alterations. A daily 30-min exposure to 1 Gz supine centrifugation at the center of mass (0.3 Gz at the head, 2 Gz at the feet) mitigated mineral density loss in the femur neck and lumbar spine; and lessened the effects of bed rest on aerobic capacity, biochemistry, and immunology. However, low statistical power could contribute to the non-significant effects that are reported. • 60-day bed rest induces decreased bone loss, muscle strength and aerobic capacity. • Bed rest also affects vascular, balance, immune, ocular, and psychology functions. • Daily 30-min of centrifugation mitigates biochemical and immunological alterations. • Centrifugation also mitigates bone loss and decreased aerobic capacity. [ABSTRACT FROM AUTHOR]

The effect of varying sinusoidal linear acceleration on perception of human motion was examined using 4 motion paradigms: off-vertical axis rotation, variable radius centrifugation, linear lateral translation, and rotation about an earth-horizontal axis. The motion profiles for each paradigm included 6 frequencies (0.01–0.6 Hz) and 5 tilt amplitudes (5°–20°). Subjects verbally reported the perceived angle of their whole-body tilt and the peak-to-peak translation of their head in space and used a joystick capable of recording 2-axis motion in the sagittal and transversal planes to indicate the phase between the perceived and actual motions. The amplitudes of perceived tilt and translation were expressed in terms of gain, i.e., the ratio of perceived tilt to equivalent tilt angle, and the ratio of perceived translation to equivalent linear displacement. Tilt perception gain decreased, whereas translation perception gain increased, with increasing frequency. During off-vertical axis rotation, the phase of tilt perception and of translation perception did not vary across stimulus frequencies. These motion paradigms elicited similar responses in roll tilt and interaural perception of translation, with differences likely due to the influence of naso-occipital linear accelerations and input to the semicircular canals that varied across motion paradigms. [ABSTRACT FROM AUTHOR]

In 2017, the study, Vision Impairment and Intracranial Pressure and Psychological Envihab Research , dubbed VaPER, was conducted in the:envihab facility in Cologne, Germany to determine whether 30 days of bed rest at 6° head down tilt in an increased ambient CO 2 (hypercapnic) environment contributes to the deterioration of ocular anatomy and to changes in musculoskeletal, nutritional, hematological, immunological, behavioral, and balance functions. A set of international standard measures were used to evaluate bone, muscle, and nutritional status, and to assess the function of the cardiovascular, sensorimotor, visual, and immune systems, and psychological state. Strict 6° head-down tilt bed rest in an elevated ambient CO 2 environment induced mild immunological dysregulation. After bed rest, selenium, urinary iodine, and folate status were lower in the subjects of the VaPER study than they were in subjects of previous bed rest studies of similar duration, which likely reflects regional differences in diet and nutritional status. Bed rest induced optic disc edema in 5 of the 11 subjects. Interestingly, the subjects who were diagnosed with optic disc edema, as determined by fundoscopy imaging, also tended to exhibit higher body temperature and overall higher levels of anxiety and insomnia during bed rest. • Strict head down tilt bed rest in an elevated ambient CO 2 environment induced mild immunological dysregulation. • After bed rest, changes in biochemistry reflected regional differences in diet and nutritional status. • Subjects who were diagnosed with optic disc edema exhibited higher body temperature and higher levels of anxiety and insomnia during bed rest. [ABSTRACT FROM AUTHOR]

We showed previously that nitrogen (N) limitation decreases Arabidopsis resistance to Erwinia amylovora (Ea). We show that decreased resistance to bacteria in low N is correlated with lower apoplastic reactive oxygen species (ROS) accumulation and lower jasmonic acid (JA) pathway expression. Consistently, pretreatment with methyl jasmonate (Me‐JA) increased the resistance of plants grown under low N. In parallel, we show that in planta titres of a nonvirulent type III secretion system (T3SS)‐deficient Ea mutant were lower than those of wildtype Ea in low N, as expected, but surprisingly not in high N. This lack of difference in high N was consistent with the low expression of the T3SS‐encoding hrp virulence genes by wildtype Ea in plants grown in high N compared to plants grown in low N. This suggests that expressing its virulence factors in planta could be a major limiting factor for Ea in the nonhost Arabidopsis. To test this hypothesis, we preincubated Ea in an inducing medium that triggers expression of hrp genes in vitro, prior to inoculation. This preincubation strongly enhanced Ea titres in planta, independently of the plant N status, and was correlated to a significant repression of JA‐dependent genes. Finally, we identify two clusters of metabolites associated with resistance or with susceptibility to Ea. Altogether, our data showed that high susceptibility of Arabidopsis to Ea, under low N or following preincubation in hrp‐inducing medium, is correlated with high expression of the Ea hrp genes in planta and low expression of the JA signalling pathway, and is correlated with the accumulation of specific metabolites. [ABSTRACT FROM AUTHOR]

Esca is one of the most widespread grapevine trunk diseases affecting vineyards. This complex disease leads to leaf alterations, wood necrosis and eventually to plant death. Esca symptoms are caused by several fungi inhabiting the xylem of host plants and degrading the wood structure. The main pathogens causing the disease are Phaeomoniella chlamydospora, Phaeoacremonium minimum, Fomitiporia mediterranea and other wood-rotting basidiomycetes. Grapevine susceptibility to esca can be predisposed by several factors, especially climate, vine age, and cultivar. An experiment was carried out (in 2015) to assess if esca expression on leaves could also be clone-dependent. Chardonnay clones 76 and 95 grown in the same plot were compared according to their developmental and physiological traits, metabolome, and foliar symptom expression. Leaves were sampled during summer on visually healthy vines as controls (C), and from asymptomatic (D-) and symptomatic (D+) shoots of esca-affected vines. Analysis of their metabolomes highlighted a clone-dependent metabolite fingerprint associated to esca expression. Opposite variations of specific metabolites were found between C and D+ leaves of both clones. The experiment was repeated (in 2018). Leaf samples could be discriminated, especially the C and D+ samples for each clone, but the differences were less marked than in the first experiment. Discriminant compounds were all different between the two experiments, and showed no opposite variations between C and D+ samples of both clones, which indicated variable metabolite responses from year to year for both clones. These results confirm that the leaf metabolite fingerprint associated to esca expression is clone-dependent, and is year-dependent in intensity and nature. [ABSTRACT FROM AUTHOR]

Space travel presents a number of environmental challenges to the central nervous system, including changes in gravitational acceleration that alter the terrestrial synergies between perception and action, galactic cosmic radiation that can damage sensitive neurons and structures, and multiple factors (isolation, confinement, altered atmosphere, and mission parameters, including distance from Earth) that can affect cognition and behavior. Travelers to Mars will be exposed to these environmental challenges for up to 3 years, and space-faring nations continue to direct vigorous research investments to help elucidate and mitigate the consequences of these long-duration exposures. This article reviews the findings of more than 50 years of space-related neuroscience research on humans and animals exposed to spaceflight or analogs of spaceflight environments, and projects the implications and the forward work necessary to ensure successful Mars missions. It also reviews fundamental neurophysiology responses that will help us understand and maintain human health and performance on Earth. [ABSTRACT FROM AUTHOR]

The perception of the horizontal and vertical distances of a visual target to an observer was investigated in parabolic flight during alternating short periods of normal gravity (1 g). microgravity (0 g), and hypergravity (1.8 g). The methods used for obtaining absolute judgments of egocentric distance included verbal reports and visually directed motion toward a memorized visual target by pulling on a rope with the arms (blind pulling). The results showed that, for all gravity levels, the verbal reports of distance judgments were accurate for targets located between 0.6 and 6.0 m. During blind pulling, subjects underestimated horizontal distances as distances increased, and this underestimation decreased in 0 g. Vertical distances for up targets were overestimated and vertical distances for down targets were underestimated in both 1 g and 1.8 g. This vertical asymmetry was absent in 0 g. The results of the present study confirm that blind pulling and verbal reports are independently influenced by gravity. The changes in distance judgments during blind pulling in 0 g compared to 1 g support the view that, during an action-based task, subjects base their perception of distance on the estimated motor effort of navigating to the perceived object. [ABSTRACT FROM AUTHOR]

BACKGROUND: The vestibulo-ocular reflex (VOR) is a basic function of the vestibular system that stabilizes gaze during head movement. Investigations on how spaceflight affects VOR gain and phase are few, and the magnitude of observed changes varies considerably and depends on the protocols used. OBJECTIVE: We investigated whether the gain and phase of the VOR in darkness and the visually assisted VOR were affected during and after spaceflight. METHODS: We measured the VOR gain and phase of 4 astronauts during and after a Space Shuttle spaceflight while the subjects voluntary oscillated their head around the yaw axis at 0.33 Hz or 1 Hz and fixed their gaze on a visual target (VVOR) or imagined this target when vision was occluded (DVOR). Eye position was recorded using electrooculography and angular velocity of the head was recorded with angular rate sensors. RESULTS: The VVOR gain at both oscillation frequencies remained near unity for all trials. DVOR gain was more variable inflight and postflight. Early inflight and immediately after the flight, DVOR gain was lower than before the flight. The phase between head and eye position was not altered by spaceflight. CONCLUSION: The decrease in DVOR gain early in the flight and after the flight reflects adaptive changes in central integration of vestibular and proprioceptive sensory inputs during active head movements. [ABSTRACT FROM AUTHOR]

Abstract Because of operational constraints, no research studies are performed during return to Earth, when astronauts perform critical operational tasks such as piloting and egressing the vehicle. The objective of this study was to interview Space Shuttle crewmembers to collect a description of post-flight perception of self-motion and self-performance from the astronauts during reentry and immediately after landing. Thirty-four Space Shuttle crewmembers were interviewed immediately after they returned from space. Transcripts of the interviews recount the crewmembers' experiences associated with head movements during reentry, during egressing the vehicle, and shortly after landing. Head movements made during reentry and immediately after return to Earth caused an increase sensation of self-motion, which affected their balance and gait. Pitch and roll head movements provoked the largest effects. Crewmembers also reported a lag in motion perception after the movement had stopped. More than one-third of crewmembers thought they would be unable to egress the Space Shuttle cabin unaided during an emergency. A clinical otoneurological examination of the crewmembers immediately after landing is needed to better understand the extent of vestibular deficits and track recovery. Highlights • 77% of crewmembers perceived larger self-motion during head movements during reentry. • 79% of crewmembers reported difficulties when standing after wheel-stop. • 36% of crewmembers felt they would be unable to egress the Space Shuttle unaided. [ABSTRACT FROM AUTHOR]

BACKGROUND: It has been proposed that individual susceptibility to motion sickness is related to the vestibulo-ocular reflex (VOR) through the activation of the velocity storage mechanism. OBJECTIVE: We investigated whether motion sickness level was related to the gain and phase of the VOR. METHODS: VOR gain and phase were measured in 214 subjects while they rotated in yaw at 0.01 Hz, 0.02 Hz, 0.04 Hz, 0.08 Hz, and 0.16 Hz in darkness, and results were compared to the severity of symptoms the subjects experienced during subsequent tests to provoke motion sickness. These tests included cross-coupled angular accelerations, sudden stops in light or in dark, off-vertical axis rotation, and parabolic flight. The subjects were grouped according to the motion sickness level reached during these tests (none, low, medium, or high). RESULTS: No correlation was found between the horizontal VOR gain and motion sickness level. However, for the subjects with high motion sickness level, the VOR phase lead was significantly lower during rotation at frequencies ranging from 0.04 Hz to 0.16 Hz (i.e. the VOR time constant was longer) than the other motion sickness groups. CONCLUSION: These results support the theory that the longer the time constant for velocity storage, the more severe the motion sickness. [ABSTRACT FROM AUTHOR]

Leaf senescence is a long developmental process important for nutrient management and for source to sink remobilization. Constituents of the mesophyll cells are progressively degraded to provide nutrients to the rest of the plant. Up to now, studies on leaf senescence have not paid much attention to the role of the different leaf tissues. In the present study, we dissected leaf laminae from the midvein to perform metabolite profiling. The laminae mesophyll cells are the source of nutrients, and in C3 plants they contain Rubisco as the most important nitrogen storage pool. Veins, rich in vasculature, are the place where all the nutrients are translocated, and sometimes interconverted, before being exported through the phloem or the xylem. The different metabolic changes we observed in laminae and midvein with ageing support the idea that the senescence programme in these two tissues is different. Important accumulations of metabolites in the midvein suggest that nutrient translocations from source leaves to sinks are mainly controlled at this level. Carbon and nitrogen long-distance molecules such as fructose, glucose, aspartate, and asparagine were more abundant in the midvein than in laminae. In contrast, sucrose, glutamate, and aspartate were more abundant in laminae. The concentrations of tricarboxylic acid (TCA) compounds were also lower in the midvein than in laminae. Since nitrogen remobilization increased under low nitrate supply, plants were grown under two nitrate concentrations. The results revealed that the senescence-related differences were mostly similar under low and high nitrate conditions except for some pathways such as the TCA cycle. [ABSTRACT FROM AUTHOR]

The root-colonizing endophytic fungus Piriformospora indica promotes the root and shoot growth of its host plants. We show that the growth promotion of Arabidopsis thaliana leaves is abolished when the seedlings are grown on media with nitrogen (N) limitation. The fungus neither stimulated the total N content nor did it promote 15NO3− uptake from agar plates to the leaves of the host under N-sufficient or N-limiting conditions. However, when the roots were co-cultivated with 15N-labelled P. indica, more labels were detected in the leaves of N-starved host plants but not in plants supplied with sufficient N. Amino acid and primary metabolite profiles, as well as the expression analyses of N metabolite transporter genes suggest that the fungus alleviates the adaptation of its host from the N limitation condition. P. indica alters the expression of transporter genes, which participate in the relocation of NO3−, NH4+ and N metabolites from the roots to the leaves under N limitation. We propose that P. indica participates in the plant's metabolomic adaptation against N limitation by delivering reduced N metabolites to the host, thus alleviating metabolic N starvation responses and reprogramming the expression of N metabolism-related genes. [ABSTRACT FROM AUTHOR]

We explored the effect of gravity on the perceived representation of the absolute distance of objects to the observers within the range from 1.5–6 m. Experiments were performed on board the CNES Airbus Zero-G during parabolic flights eliciting repeated exposures to short periods of microgravity (0 g), hypergravity (1.8 g), and normal gravity (1 g). Two methods for obtaining estimates of perceived egocentric distance were used: verbal reports and visually directed motion toward a memorized visual target. For the latter method, because normal walking is not possible in 0 g, blindfolded subjects translated toward the visual target by pulling on a rope with their arms. The results showed that distance estimates using both verbal reports and blind pulling were significantly different between normal gravity, microgravity, and hypergravity. Compared to the 1 g measurements, the estimates of perceived distance using blind pulling were shorter for all distances in 1.8 g, whereas in 0 g they were longer for distances up to 4 m and shorter for distances beyond. These findings suggest that gravity plays a role in both the sensorimotor system and the perceptual/cognitive system for estimating egocentric distance. [ABSTRACT FROM AUTHOR]

The chemical composition of plant surfaces plays a role in selection of host plants by herbivorous insects. Once the insect reaches the plant, these cues determine host acceptance. Laboratory studies have shown that the stem borer Busseola fusca (Lepidoptera: Noctuidae), an important pest of sorghum and maize in sub-Saharan Africa, is able to differentiate between host and non-host plant species. However, no information is available on the cues used by this insect to seek and accept the host plant. Thus, the role of surface phytochemical stimuli on host selection and oviposition by B. fusca was studied in the laboratory using two host plants, sorghum, Sorghum bicolor, and maize, Zea mays, and one non-host plant, Napier grass, Pennisetum purpureum. The numbers of eggs and egg masses deposited on the three plant species were compared first under no-choice and choice conditions. In both cases, more eggs and egg masses were laid on maize and sorghum than on the non-host. Artificial surrogate stems treated with a water or chloroform surface extract of each plant were then compared with surrogate stems treated with, respectively, water or chloroform as controls, under similar conditions. Surrogate stems treated with plant water extracts did not show an increase in oviposition when compared to controls, indicating that the major compounds in these extracts, i.e., simple sugars and free amino acids, are not significantly responsible for the oviposition preference. By contrast, a chloroform extract of sorghum enhanced oviposition on the surrogate stems compared to the control, while those of maize and Napier grass showed no significant effects. Analysis of the chloroform extract of sorghum showed higher amounts of α-amyrin, ß-amyrin, and n-nonacosane compared to those of maize and Napier grass. A blend of the three chemicals significantly increased oviposition compared to the chloroform-treated control, indicating that these compounds are part of the surface chemical signature of the plant responsible for host recognition and oviposition by B. fusca. [ABSTRACT FROM AUTHOR]

Two characteristics of otolith-ocular responses – linear vestibulo-ocular reflex and vergence – were examined during constant velocity off-vertical axis rotation (OVAR) in the dark. Sixteen subjects were rotated about their longitudinal axis when tilted 30° relative to the direction of gravity. Rotational velocities were 36 and 288/s corresponding to frequencies of 0.1 and 0.8 Hz, respectively. Subjects were asked to imagine stationary targets located at 0.5 m, 1 m, and 2 m in the straight-ahead direction. Binocular eye movements were recorded in the dark using infrared videography. The modulation of horizontal slow phase velocity during OVAR was larger at 0.8 Hz than at 0.1 Hz, and the modulation at the high frequency was larger for the near target than for the mid and far targets. These characteristics confirm that the horizontal slow phase velocity during yaw OVAR represents a translational otolith-ocular reflex in response to acceleration along the inter-aural axis that is dependent on imagined fixation distance. [ABSTRACT FROM AUTHOR]

NAD-dependent glutamate dehydrogenase (NAD-GDH) of higher plants has a central position at the interface between carbon and nitrogen metabolism due to its ability to carry out the deamination of glutamate. In order to obtain a better understanding of the physiological function of NAD-GDH under salt stress conditions, transgenic tobacco (Nicotiana tabacum L.) plants that overexpress two genes from Nicotiana plumbaginifolia individually (GDHA and GDHB) or simultaneously (GDHA/B) were grown in the presence of 50 mM NaCl. In the different GDH overexpressors, the NaCl treatment induced an additional increase in GDH enzyme activity, indicating that a post-transcriptional mechanism regulates the final enzyme activity under salt stress conditions. A greater shoot and root biomass production was observed in the three types of GDH overexpressors following growth in 50 mM NaCl, when compared with the untransformed plants subjected to the same salinity stress. Changes in metabolites representative of the plant carbon and nitrogen status were also observed. They were mainly characterized by an increased amount of starch present in the leaves of the GDH overexpressors as compared with the wild type when plants were grown in 50mM NaCl. Metabolomic analysis revealed that overexpressing the two genes GDHA and GDHB, individually or simultaneously, induced a differential accumulation of several carbon- and nitrogen-containing molecules involved in a variety of metabolic, developmental and stress-responsive processes. An accumulation of digalactosylglycerol, erythronate and porphyrin was found in the GDHA, GDHB and GDHA/B overexpressors, suggesting that these molecules could contribute to the improved performance of the transgenic plants under salinity stress conditions. [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate depth perception in astronauts during and after spaceflight by studying their sensitivity to reversible perspective figures in which two-dimensional images could elicit two possible depth representations. Other ambiguous figures that did not give rise to a perception of illusory depth were used as controls. Six astronauts and 14 subjects were tested in the laboratory during three sessions for evaluating the variability of their responses in normal gravity. The six astronauts were then tested during four sessions while on board the International Space Station for 5–6 months. They were finally tested immediately after return to Earth and up to one week later. The reaction time decreased throughout the sessions, thus indicating a learning effect. However, the time to first percept reversal and the number of reversals were not different in orbit and after the flight compared to before the flight. On Earth, when watching depth-ambiguous perspective figures, all subjects reported seeing one three-dimensional interpretation more often than the other, i.e. a ratio of about 70–30%. In weightlessness this asymmetry gradually disappeared and after 3 months in orbit both interpretations were seen for the same duration. These results indicate that the perception of “illusory” depth is altered in astronauts during spaceflight. This increased depth ambiguity is attributed to the lack of the gravitational reference and the eye-ground elevation for interpreting perspective depth cues. [ABSTRACT FROM AUTHOR]

The central nervous system must resolve the ambiguity of inertial motion sensory cues in order to derive an accurate representation of spatial orientation. Adaptive changes during spaceflight in how the brain integrates vestibular cues with other sensory information can lead to impaired movement coordination, vertigo, spatial disorientation, and perceptual illusions after return to Earth. The purpose of this study was to compare tilt and translation motion perception in astronauts before and after returning from spaceflight. We hypothesized that these stimuli would be the most ambiguous in the low-frequency range (i.e., at about 0.3 Hz) where the linear acceleration can be interpreted either as a translation or as a tilt relative to gravity. Verbal reports were obtained in eleven astronauts tested using a motion-based tilt-translation device and a variable radius centrifuge before and after flying for two weeks on board the Space Shuttle. Consistent with previous studies, roll tilt perception was overestimated shortly after spaceflight and then recovered with 1–2 days. During dynamic linear acceleration (0.15–0.6 Hz, ±1.7 m/s2) perception of translation was also overestimated immediately after flight. Recovery to baseline was observed after 2 days for lateral translation and 8 days for fore–aft translation. These results suggest that there was a shift in the frequency dynamic of tilt-translation motion perception after adaptation to weightlessness. These results have implications for manual control during landing of a space vehicle after exposure to microgravity, as it will be the case for human asteroid and Mars missions. [ABSTRACT FROM AUTHOR]

Abstract: This paper explores the evolution of human spaceflight by examining the space programs of the United States, Russia, including the former Soviet Union, and China. A simple analysis of the numbers of humans who have flown into space, the durations of the missions flown, and the accumulated flight time of the individuals reveals that spaceflight is decidedly male-dominated and that approximately one out of six individuals flown was a non-career astronaut. In addition, 31 individuals have accumulated long-duration flight experience equivalent to a round trip to Mars. An examination of the evolution of spacecraft that have made these missions possible indicates that the time to accomplish the first four to five flights of a new human space vehicle has increased from less than one year to nearly 10 years. [Copyright &y& Elsevier]

When an observer lifts two objects with the same weight but different sizes, the smaller object is consistently reported to feel heavier than the larger object even after repeated trials. Here we explored the effect of reduced and increased gravity on this perceptual size-mass illusion. Experiments were performed on board the CNES Airbus A300 Zero-G during parabolic flights eliciting repeated exposures to short periods of zero g, 0.16 g, 0.38 g, one g, and 1.8 g. Subjects were asked to assess perceived heaviness by actively oscillating objects with various sizes and masses. The results showed that a perceptual size-mass illusion was clearly present at all gravity levels. During the oscillations, the peak arm acceleration varied as a function of the gravity level, irrespective of the mass and size of the objects. In other words we did not observe a sensorimotor size-mass illusion. These findings confirm dissociation between the sensorimotor and perceptual systems for determining object mass. In addition, they suggest that astronauts on the Moon or Mars with the eyes closed will be able to accurately determine the relative difference in mass between objects. [ABSTRACT FROM AUTHOR]

Autophagy is a fundamental process in the plant life story, playing a key role in immunity, senescence, nutrient recycling, and adaptation to the environment. Transcriptomics and metabolomics of the rosette leaves of Arabidopsis thaliana autophagy mutants (atg) show that autophagy is essential for cell homeostasis and stress responses and that several metabolic pathways are affected. Depletion of hexoses, quercetins, and anthocyanins parallel the overaccumulation of several amino acids and related compounds, such as glutamate, methionine, glutathione, pipecolate, and 2-aminoadipate. Transcriptomic data show that the pathways for glutathione, methionine, raffinose, galacturonate, and anthocyanin are perturbed. Anthocyanin depletion in atg mutants, which was previously reported as a possible defect in flavonoid trafficking to the vacuole, appears due to the downregulation of the master genes encoding the enzymes and regulatory proteins involved in flavonoid biosynthesis. Overexpression of the PRODUCTION OF ANTHOCYANIN PIGMENT1 transcription factor restores anthocyanin accumulation in vacuoles of atg mutants. Transcriptome analyses reveal connections between autophagy and (1) salicylic acid biosynthesis and response, (2) cytokinin perception, (3) oxidative stress and plant defense, and possible interactions between autophagy and the COP9 signalosome machinery. The metabolic and transcriptomic signatures identified for the autophagy mutants are discussed and show consistencies with the observed phenotypes. [ABSTRACT FROM AUTHOR]

BACKGROUND: Artificial gravity using short-radius centrifugation has been proposed as an integrative countermeasure during spaceflight.OBJECTIVE: To determine the rotation parameters of a short-radius centrifuge so that subjects rotating in the dark would feel as if they were standing upright. METHODS: Twelve subjects were lying supine in a nacelle on a 2.8 m-radius centrifuge with their head closer to the axis of rotation and their feet pointing radially outwards. Subjects verbally reported body orientation for 26 combinations of centrifuge rotation rate and nacelle pitch tilt. ECG and respiratory responses were also recorded. RESULTS: Five subjects felt like they were vertical when centrifugation elicited 1 g at their center of mass along their body longitudinal axis, whereas seven subjects felt they were vertical when they experienced about 1 g at ear level, regardless of the nacelle tilt angle. Heart rate variability varied with the subjects' perception of verticality. CONCLUSIONS: These results suggest that one group of subject was relying principally on the otolith organs for the perception of verticality, whereas the other group was also relying on extravestibular somatosensory receptors. The crewmember's perception of verticality might be a factor to take into account for the prescription for artificial gravity during spaceflight. [ABSTRACT FROM AUTHOR]

Exposure to microgravity during spaceflight is known to elicit orientation illusions, errors in sensory localization, postural imbalance, changes in vestibulo-spinal and vestibulo-ocular reflexes, and space motion sickness. The objective of this experiment was to investigate whether an alteration in cognitive visual-spatial processing, such as the perception of distance and size of objects, is also taking place during prolonged exposure to microgravity. Our results show that astronauts on board the International Space Station exhibit biases in the perception of their environment. Objects' heights and depths were perceived as taller and shallower, respectively, and distances were generally underestimated in orbit compared to Earth. These changes may occur because the perspective cues for depth are less salient in microgravity or the eye-height scaling of size is different when an observer is not standing on the ground. This finding has operational implications for human space exploration missions. [ABSTRACT FROM AUTHOR]

Abstract: Preliminary results of an ongoing study examining the effects of space flight on astronauts' motion perception induced by independent tilt and translation motions are presented. This experiment used a sled and a variable radius centrifuge that translated the subjects forward-backward or laterally, and simultaneously tilted them in pitch or roll, respectively. Tests were performed on the ground prior to and immediately after landing. The astronauts were asked to report about their perceived motion in response to different combinations of body tilt and translation in darkness. Their ability to manually control their own orientation was also evaluated using a joystick with which they nulled out the perceived tilt while the sled and centrifuge were in motion. Preliminary results confirm that the magnitude of perceived tilt increased during static tilt in roll after space flight. A deterioration in the crewmember to control tilt using non-visual inertial cues was also observed post-flight. However, the use of a tactile prosthesis indicating the direction of down on the subject's trunk improved manual control performance both before and after space flight. [Copyright &y& Elsevier]

Highlights: [•] Time interval between velocity steps influences vestibular habituation of nystagmus. [•] VOR habituates only when steps are delivered after the primary phase nystagmus. [•] Perception of self-rotation habituates irrespectively of stimulus interval. [Copyright &y& Elsevier]

Experiments performed in orbit on the central nervous system have focused on the control of posture, eye movements, spatial orientation, as well as cognitive processes, such as three-dimensional visual perception and mental representation of space. Brain activity has also been recorded during and immediately after space flight for evaluating the changes in brain structure activation during tasks involving perception, attention, memory, decision, and action. Recent ground-based studies brought evidence that the inputs from the neurovestibular system also participate in orthostatic intolerance. It is, therefore, important to revisit the flight data of neuroscience studies in the light of new models of integrative physiology. The outcomes of this exercise will increase our knowledge on the adaptation of body functions to changing gravitational environment, vestibular disorders, aging, and our approach towards more effective countermeasures during human space flight and planetary exploration. [ABSTRACT FROM AUTHOR]

Experiments performed in orbit on the central nervous system have focused on the control of posture, eye movements, spatial orientation, as well as cognitive processes, such as three-dimensional visual perception and mental representation of space. Brain activity has also been recorded during and immediately after space flight for evaluating the changes in brain structure activation during tasks involving perception, attention, memory, decision, and action. Recent ground-based studies brought evidence that the inputs from the neurovestibular system also participate in orthostatic intolerance. It is, therefore, important to revisit the flight data of neuroscience studies in the light of new models of integrative physiology. The outcomes of this exercise will increase our knowledge on the adaptation of body functions to changing gravitational environment, vestibular disorders, aging, and our approach towards more effective countermeasures during human space flight and planetary exploration. [ABSTRACT FROM AUTHOR]

Constant velocity off-vertical axis rotation (OVAR) provides dynamic linear acceleration stimuli that can be used to assess otolith function. Eight astronauts were rotated in darkness about their longitudinal axis 20° off vertical at low (0.125 Hz) and high (0.5 Hz) frequencies and their responses were compared before and after spaceflight. Eye movements were recorded using infrared videography and perceived motion was evaluated using a joystick with four degrees of freedom - pitch and roll tilt, front-back and lateral translation. Low-frequency OVAR generates tilt otolith-induced responses - modulation of ocular counter-roll and counter-pitch with perceived conical motion path - whereas high-frequency OVAR generates translational otolith-induced responses - modulation of horizontal and vergence slow phase velocity with perceived cylindrical motion path. While there were transient changes in the amplitude of the translational ocular responses on landing day, there were no major changes in the tilt ocular reflexes after adaptation to weightlessness. However, there was an increase in sensitivity to motion perception after spaceflight. Direct comparisons of pre- and postflight stimuli suggested that OVAR on landing day was less provocative of motion sickness than before spaceflight. These results confirm that some otolith reflexes elicited during passive motion may not be altered by short-duration spaceflight - or may readapt very quickly - and that the resolution of sensory conflict associated with postflight recovery involves higher-order neural processes. [ABSTRACT FROM AUTHOR]

Abstract: Although the mechanisms of neural adaptation to weightlessness and re-adaptation to Earth-gravity have received a lot of attention since the first human space flight, there is as yet little knowledge about how spatial orientation is affected by partial gravity, such as lunar gravity of 0.16g or Martian gravity of 0.38g. Up to now twelve astronauts have spent a cumulated time of approximately 80h on the lunar surface, but no psychophysical experiments were conducted to investigate their perception of verticality. We investigated how the subjective vertical (SV) was affected by reduced gravity levels during the first European Parabolic Flight Campaign of Partial Gravity. In normal and hypergravity, subjects accurately aligned their SV with the gravitational vertical. However, when gravity was below a certain threshold, subjects aligned their SV with their body longitudinal axis. The value of the threshold varied considerably between subjects, ranging from 0.03 to 0.57g. Despite the small number of subjects, there was a significant positive correlation of the threshold with subject age, which calls for further investigation. [Copyright &y& Elsevier]

Abstract: This experiment investigated whether the perception of depth-reversible figures is altered when the observer is in microgravity or hypergravity. A set of five bi-stable ambiguous figures was presented to ten participants in 1g, 0g, and 1.8g during parabolic flight. The figures included static images such as the Necker cube; kinetic depth displays such as a moving plaid and a sphere cluster of moving dots appearing to rotate in one of two directions; and a silhouette photograph. For each stimulus figure, subjects reported which of the two possible perceptual configurations they saw first and then continuously indicated when perceptual reversals occurred for durations ranging from 20 to 30s. The same first percept was reported in 1g, 0g, and 1.8g. The time delay for the first reversal between the two possible image interpretations was longer and the number of reversals was fewer in 0g as compared to 1g for four of the five figures. The opposite effects were seen when going from 0g to 1.8g. These findings confirm that, consistent with a multisensory approach to three-dimensional form perception, gravity has a clear effect on the interpretation of depth-based stimuli and this gravity-based component interferes with visual perception stability. [Copyright &y& Elsevier]

The development of therapeutic recombinant antibodies involves accurate characterization of immunoglobulin variable light (VL) and heavy (VH) chains. However, it has been reported that the use of subgroup or isotype-specific primers for the amplification of monoclonal antibody (mAb) variable domains introduces heterogeneities within the variable domains, or amplifies aberrant productive Ig domains. To address these issues, we have combined the rapid amplification of cDNA ends PCR (RACE-PCR) for the full-length VL and VH amplification, with peptide mass fingerprinting of the corresponding Ig chain. Using this strategy, we amplified full-length cDNA chains of SAF34 and SAF32, two potential therapeutic mAbs against neurodegenerative diseases directed to the prion protein (PrP). We report an unambiguous correlation between hybridoma cDNA sequences and protein fingerprints of the variable domains of each mAb, indicating the discrimination between mutated, pseudo-genes and functional Ig genes. As a proof of principle for this dual strategy of full-length PCR amplification of variable domains and their characterization by MALDI-TOF, we show that the corresponding scFvs recognize the native PrP and retain full capacity to bind to human PrP, as does the parental mAb. This finding addresses the need for reliable light and heavy chain characterization, a key factor for humanization of mouse antibodies and for its use in passive immunotherapy applications. [ABSTRACT FROM PUBLISHER]

Abstract: Four patients following unilateral vestibular loss and four patients complaining of otolith-dependent vertigo were tested during eccentric yaw rotation generating 1× g centripetal acceleration directed along the interaural axis. Perception of body tilt in roll and in pitch was recorded in darkness using a somatosensory plate that the subjects maintained parallel to the perceived horizon. Ocular torsion was recorded by a video camera. Unilateral vestibular-defective patients underestimated the magnitude of the roll tilt and had a smaller torsion when the centrifugal force was towards the operated ear compared to the intact ear and healthy subjects. Patients with otolithic-dependent vertigo overestimated the magnitude of roll tilt in both directions of eccentric rotation relative to healthy subjects, and their ocular torsion was smaller than in healthy subjects. Eccentric rotation is a promising tool for the evaluation of vestibular dysfunction in patients. Eye torsion and perception of tilt during this stimulation are objective and subjective measurements, which could be used to determine alterations in spatial processing in the CNS. [Copyright &y& Elsevier]

Abstract: Humans have mental representation of their environment based on sensory information and experience. A series of experiments has been designed to allow the identification of disturbances in the mental representation of three-dimensional space during space flight as a consequence of the absence of the gravitational frame of reference. This NASA/ESA-funded research effort includes motor tests complemented by psychophysics measurements, designed to distinguish the effects of cognitive versus perceptual-motor changes due to microgravity exposure. Preliminary results have been obtained during the microgravity phase of parabolic flight. These results indicate that the vertical height of handwritten characters and drawn objects is reduced in microgravity compared to normal gravity, suggesting that the mental representation of the height of objects and the environment change during short-term microgravity. Identifying lasting abnormalities in the mental representation of spatial cues will establish the scientific and technical foundation for development of preflight and in-flight training and rehabilitative schemes, enhancing astronaut performance of perceptual-motor tasks, for example, interaction with robotic systems during exploration-class missions. [Copyright &y& Elsevier]

Abstract: In a classic demonstration, Ernst Mach showed that the same figure could be perceived as a square or as a diamond depending on the orientation of the subject relative to gravity. Such phenomenon is based on the use of a geocentric reference frame for object perception. If the central nervous system perceives an object with respect to the gravitationally defined vertical, what will happen if this reference frame is removed? We investigated the Mach phenomenon in subjects placed in short-term microgravity during parabolic flight. Subjects were presented with a square with a corner pointing upwards, and asked whether they perceived it as a diamond or square with the head upright or tilted 45° in roll both in normal gravity and when free-floating in microgravity during parabolic flight. The addition of a rectangular frame around the figure was also investigated. In contrast to the normal gravity condition, with the head tilted the subjects still perceived a diamond figure in microgravity, indicating that they had switched from a geocentric to an egocentric reference frame. Also in contrast to the normal gravity condition, adding a rectangular frame around the figure did not significantly change the perception of the object in microgravity, suggesting that an intrinsic reference determined by the axis of elongation or symmetry of the object does not easily override an egocentric reference frame like it does for a geocentric reference frame. [Copyright &y& Elsevier]

Abstract: This investigation examined depth and distance perception of 3-D objects presented in stereoscopic vision. The results obtained in normal gravity and microgravity in parabolic flight were compared. This experiment was performed during an ESA campaign of parabolic flight on the Airbus A300 Zero-G on six free-floating subjects. The subjects were first presented with a 3-D cube and were instructed to adjust one dimension (width, length, or height) of the cube so that all three dimensions had the same perceived length. The subjects perceived a normal cube to be taller, thinner, and shallower in microgravity compared to normal gravity. Consequently, when they adjusted its dimension so that it looked “normal” to them, they made it shorter, wider, and deeper. A simple distance perception test confirmed that the subjects underestimated the distance of objects in the depth plane in microgravity. These results confirm a role of the gravitational reference in the 3-D perception of the objects and the environment. [Copyright &y& Elsevier]

Abstract: Many experts believe that artificial gravity will be required for an interplanetary mission. However, despite its attractiveness as an efficient, multi-system countermeasure and its potential for simplifying operational activities, much still needs to be learned regarding the human response to rotating environments before artificial gravity can be successfully implemented. The European Space Agency (ESA) Topical Team on Artificial Gravity recommended a comprehensive program to determine the gravity threshold required to reverse or prevent the detrimental effects of microgravity and to evaluate the effects of centrifugation on various physiological functions. Part of the required research can be accomplished using animal models on a dedicated centrifuge in low Earth orbit. Studies of human responses to centrifugation could be performed during ambulatory, short- and long-duration bed rest, and in-flight studies. Artificial-gravity scenarios should not be a priori discarded in Moon and Mars mission designs. One major step is to determine the relationship between the artificial gravity dose level, duration, and frequency and the physiological responses of the major body functions affected by spaceflight. Once its regime characteristics are defined and a dose–response curve is established, artificial gravity should serve as the standard against which all other countermeasure candidates are evaluated, first on Earth and then in space. [Copyright &y& Elsevier]

In humans, habituation of vestibulo-ocular reflex (VOR) by repeated caloric or rotational stimulation has been well documented. However, less attention has been directed to the effect of habituation on the sensation of self-rotation and little is known about the retention duration of vestibular habituation. To investigate these characteristics, subjects were exposed to ten sessions of angular velocity steps in yaw, with a chair rotating either alternatively in both CW and CCW directions (bidirectional protocol) or always in the same direction (unidirectional protocol), i.e., CW or CCW. The retention of habituation of VOR and sensation of rotation induced by both protocols was studied for a period up to 8 months following the end of the habituation protocols. There was a progressive decline in the VOR peak slow phase velocity and time constant throughout the sessions during both protocols. These parameters then followed an exponential recovery with a time constant of about 1 month. The duration of the sensation of rotation also habituated during repeated angular velocity steps, but it was shorter for both directions of stimulation, including after the unidirectional protocol. Sinusoidal VOR gain was not affected by vestibular habituation to velocity steps, but sinusoidal VOR phase showed an increase in phase lead at 0.02 and 0.04 Hz, which also returned to baseline values within about 1 month. We conclude that vestibular habituation is a long-lasting phenomenon. These results may be helpful for designing and scheduling the protocols for drug studies using crossover design, rehabilitation of balance disorder patients, and for the application of intermittent artificial gravity during space missions. [ABSTRACT FROM AUTHOR]

Ocular counter-rolling (OCR) induced by whole body tilt in roll has been explored after spaceflight as an indicator of the adaptation of the otolith function to microgravity. It has been claimed that the overall pattern of OCR responses during static body tilt after spaceflight is indicative of a decreased role of the otolith function, but the results of these studies have not been consistent, mostly due to large variations in the OCR within and across individuals. By contrast with static head tilt, off-vertical axis rotation (OVAR) presents the advantage of generating a sinusoidal modulation of OCR, allowing averaged measurements over several cycles, thus improving measurement accuracy. Accordingly, OCR and the sense of roll tilt were evaluated in seven astronauts before and after spaceflight during OVAR at 45°/s in darkness at two angles of tilt (10° and 20°). There was no significant difference in OCR during OVAR immediately after landing compared to preflight. However, the amplitude of the perceived roll tilt during OVAR was significantly larger immediately postflight, and then returned to control values in the following days. Since the OCR response is predominantly attributed to the shearing force exerted on the utricular macula, the absence of change in OCR postflight suggests that the peripheral otolith organs function normally after short-term spaceflight. However, the increased sense of roll tilt indicates an adaptation in the central processing of gravitational input, presumably related to a re-weighting of the internal representation of gravitational vertical as a result of adaptation to microgravity. [ABSTRACT FROM AUTHOR]

Pitch head-and-trunk movements during constant velocity rotation are a provocative vestibular stimulus that produces vertigo and nausea. When exposed to this stimulus repeatedly, motion sickness symptoms diminish as the subjects habituate. Acetylleucine is a drug that is used to treat acute vestibular vertigo. In this study, we wanted to ascertain whether this drug (a) lessened motion sickness or delayed habituation; (b) accelerated the recovery following habituation; and (c) whether changes in the subjective vertical accompanied habituation. Twenty subjects were administered acetylleucine or placebo in a double-blind study during a five-day vestibular training. Horizontal vestibulo-ocular reflex, optokinetic nystagmus, smooth pursuit, and subjective visual vertical were evaluated before, during, and up to two months after the vestibular training. Based on Graybiel's diagnostic criteria, motion sickness decreased steadily in each vestibular training session, and there was no difference between the scores in the acetylleucine and placebo groups. Post-rotatory nystagmus peak velocity and time constant also declined in both groups at the same rate. Thus, acetylleucine neither reduced the nausea associated with this provocative stimulus, nor hastened the acquisition or retention of vestibular habituation of motion sickness and nystagmus. There was no difference in optokinetic nystagmus and smooth pursuit between the acetylleucine and placebo groups. However, subjects showed larger error in the subjective visual vertical after habituation, which indicates that spatial orientation is also affected by vestibular training. [ABSTRACT FROM AUTHOR]

Abstract: It has been proposed that an internal representation of body vertical has a prominent role in spatial orientation. This investigation investigated the ability of human subjects to accurately locate their longitudinal body axis (an imaginary straight body midline running from head to toes) while free-floating in microgravity. Subjects were tested in-flight, as well as on ground in normal gravity in both the upright and supine orientations to provide baseline measurements. The subjects wore a goggle device and were in total darkness. They used knobs to rotate two luminous lines until they were parallel to the subjective direction of their longitudinal body axis, in the roll (right/left) and the pitch (forward/backward) planes. Results showed that the error between the perceived and the objective direction of the longitudinal body axis was significantly larger in microgravity than in normal gravity. This error in this egocentric frame of reference is presumably due to the absence of somatosensory cues when free-floating. Mechanical pressure on the chest using an airbag reduced the error in perception of the longitudinal body axis in microgravity, thus improving spatial orientation. [Copyright &y& Elsevier]

Objective: There are a number of physiological changes which occur in astronauts in both short- and long-duration space missions, including nausea and spatial disorientation, orthostatic hypotension, muscle atrophy, bone loss, increased cancer risk from space radiation, and many others. This review examines the procedures and methods, also called countermeasures, used to moderate these changes. Data sources: The information in this paper is taken from a review of articles and book chapters (Source: PubMed and MEDLINE, years covered 1995-2005). Conclusions: The countermeasures currently adopted to counteract the effects of microgravity conditions on board space missions aim at stimulating a particular physiological system: the treadmill, the cycle ergometer and the interim resistive exercise device primarily for muscles and bones, intermittent venous pooling and fluid loading for cardiovascular responses, and pharmacological manipulations for space motion sickness. However, all have only limited success. Indeed, despite extensive in-flight exercise, most astronauts experience difficulties in standing, walking and getting oriented for several days after landing. This poses a serious problem in case of an emergency landing on Earth or a landing on Mars after a long-duration spaceflight. Studies are being conducted for the search of more effective countermeasures that address all physiological systems across the board, such as artificial gravity generated by short-arm centrifugation. [ABSTRACT FROM AUTHOR]

Abstract: The occurrence of geometric orientation illusions and the perception of ambiguous figures were analyzed in 24 subjects during static body tilt relative to gravity on Earth. Results showed that illusions such as the Rock''s diamond/square, the Ponzo illusion, and orientation contrast illusions occurred less frequently, and that depth reversal of ambiguous figures took more time when subjects were lying on their side or supine compared to upright, thus suggesting that the gravitational reference plays a significant role in these “visual” illusions. The structure of images, our representation of the environment, and orientation relative to gravity are all integral parts in interpreting visual images. In a weightless environment where no gravitational reference can be used, it is expected that similar alterations in visual perception will occur. [Copyright &y& Elsevier]

The human vestibulo-ocular reflex (VOR) was investigated during active head movements in yaw while subjects were asked to view targets located at 20, 30, 40, 60, 90, and 120 cm distance aligned with eye level. Binocular video cameras were used to study conjugate eye movements and binocular convergence. Perceived target distance was determined during head oscillation by having the subjects move a cursor to the remembered position of the previously seen targets. The changes in VOR gain with viewing distance were found to be more closely related to perceived target distance than to actual target distance or fixation distance. This result suggests that the adjustment of VOR gain with viewing distance is under stronger cognitive control than would be expected of a simple motor reflex. [Copyright &y& Elsevier]

In this paper we review space flight experiments performed by our laboratory. Rhesus monkeys were tested before and after 12 days in orbit on COSMOS flights 2044 (1989) and 2229 (1992–1993). There was a long-lasting decrease in post-flight ocular counter-rolling (70%) and vergence (50%) during off-vertical axis rotation. In one animal, the orientation of optokinetic after-nystagmus shifted by 28° from the spatial vertical towards the body vertical early post-flight. Otolith-ocular and perceptual responses were also studied in four astronauts on the 17-day Neurolab shuttle mission (STS-90) in 1998. Ocular counter-rolling was unchanged in response to 1-g and 0.5-g Gy centrifugation during and after flight and to post-flight static roll tilts relative to pre-flight values. Orientation of the optokinetic nystagmus eye velocity axis to gravito-inertial acceleration (GIA) during centrifugation was also unaltered by exposure to microgravity. Perceptual orientation to the GIA was maintained in-flight, and subjects did not report sensation of translation during constant velocity centrifugation. These studies suggest that percepts and ocular responses to tilt are determined by sensing the body vertical relative to the GIA. The findings also raise the possibility that 'artificial gravity' during the Neurolab flight counteracted adaptation of these otolith-ocular responses. [ABSTRACT FROM AUTHOR]

Prolonged microgravity during orbital flight is a unique way to modify the otolith inputs and to determine the extent of their contribution to the vertical vestibulo-ocular reflex (VOR) and optokinetic nystagmus (OKN). This paper reviews the data collected on 10 astronauts during several space missions and focuses on the changes in the up-down asymmetry. Both the OKN elicited by vertical visual stimulation and the active VOR elicited by voluntary pitch head movements showed an asymmetry before flight, with upward slow phase velocity higher than downward slow phase velocity. Early in-flight, this asymmetry was inverted, and a symmetry of both responses was later observed. An upward shift in the vertical mean eye position in both OKN and VOR suggests that these effects may be related to otolith-dependent changes in eye position which, in themselves, affect slow phase eye velocity. [ABSTRACT FROM AUTHOR]

SUMMARY: The SID2 (SA INDUCTION‐DEFICIENT2) gene that encodes ICS1 (isochorismate synthase), plays a central role in salicylic acid biosynthesis in Arabidopsis. The sid2 and NahG (encoding a bacterial SA hydroxylase) overexpressing mutants (NahG‐OE) have currently been shown to outperform wild type, presenting delayed leaf senescence, higher plant biomass and better seed yield. When grown under sulfate‐limited conditions (low‐S), sid2 mutants exhibited early leaf yellowing compared to the NahG‐OE, the npr1 mutant affected in SA signaling pathway, and WT. This indicated that the hypersensitivity of sid2 to sulfate limitation was independent of the canonical npr1 SA‐signaling pathway. Transcriptomic and proteomic analyses revealed that major changes occurred in sid2 when cultivated under low‐S, changes that were in good accordance with early senescence phenotype and showed the exacerbation of stress responses. The sid2 mutants displayed a lower sulfate uptake capacity when cultivated under low‐S and lower S concentrations in their rosettes. Higher glutathione concentrations in sid2 rosettes under low‐S were in good accordance with the higher abundance of proteins involved in glutathione and ascorbate redox metabolism. Amino acid and lipid metabolisms were also strongly modified in sid2 under low‐S. Depletion of total fatty acids in sid2 under low‐S was consistent with the fact that S‐metabolism plays a central role in lipid synthesis. Altogether, our results show that functional ICS1 is important for plants to cope with S limiting conditions. [ABSTRACT FROM AUTHOR]

DspA/E is a type three effector injected by the pathogenic bacterium Erwinia amylovora inside plant cells. In non-host Arabidopsis thaliana, DspA/E inhibits seed germination, root growth, de novo protein synthesis and triggers localized cell death. To better understand the mechanisms involved, we performed EMS mutagenesis on a transgenic line, 13-1-2, containing an inducible dspA/E gene. We identified three suppressor mutants, two of which belonged to the same complementation group. Both were resistant to the toxic effects of DspA/E. Metabolome analysis showed that the 13-1-2 line was depleted in metabolites of the TCA cycle and accumulated metabolites associated with cell death and defense. TCA cycle and cell-death associated metabolite levels were respectively increased and reduced in both suppressor mutants compared to the 13-1-2 line. Whole genome sequencing indicated that both suppressor mutants displayed missense mutations in conserved residues of Glycolate oxidase 2 (GOX2), a photorespiratory enzyme that we confirmed to be localized in the peroxisome. Leaf GOX activity increased in leaves infected with E. amylovora in a DspA/E-dependent manner. Moreover, the gox2-2 KO mutant was more sensitive to E. amylovora infection and displayed reduced JA-signaling. Our results point to a role for glycolate oxidase in type II non-host resistance and to the importance of central metabolic functions in controlling growth/defense balance. [ABSTRACT FROM AUTHOR]

Esca is a complex grapevine trunk disease caused by wood‐rotting ascomycetes and basidiomycetes and leading to several foliar and wood symptoms. Given that the esca expression can be influenced by several environmental, physiological, and genetic factors, foliar symptoms are inconsistent in incidence and prevalence and may appear 1 year but not the following. We have previously reported a clone‐dependent expression of the disease in cv Chardonnay. Owing to metabolome analysis, we could discriminate the metabolite fingerprint of green leaves collected on diseased vines of clones 76 and 95. These clone‐dependent fingerprints were year‐dependent in intensity and nature. The present work was conducted to determine if the clone‐dependent disease expression observed is specific to Chardonnay or if it also occurs in another cultivar. A plot located in the Jura vineyard (France) and planted with both 1004 and 1026 clones of Trousseau, a cultivar highly susceptible to esca, was thus selected and studied during 2017 and 2018. A year‐dependent variation of the symptoms expression was first observed and a possible relationship with rainfall is hypothesized and discussed. Moreover, a higher percentage of the clone 1026 vines expressed disease, compared to the 1004 ones, suggesting the higher susceptibility of this clone. Finally, metabolomic analyses of the remaining green leaves (i.e, without symptom expression) of partial esca‐apoplectic vines allowed us to confirm a clone‐dependent metabolic response to the disease. The metabolite fingerprints obtained differed in nature and intensity to those previously reported for Chardonnay and also between years. [ABSTRACT FROM AUTHOR]

Plants have fundamental dependences on nitrogen and sulfur and frequently have to cope with chronic limitations when their supply is sub-optimal. This study aimed at characterizing the metabolomic, proteomic, and transcriptomic changes occurring in Arabidopsis leaves under chronic nitrate (Low-N) and chronic sulfate (Low-S) limitations in order to compare their effects, determine interconnections, and examine strategies of adaptation. Metabolite profiling globally revealed opposite effects of Low-S and Low-N on carbohydrate and amino acid accumulations, whilst proteomic data showed that both treatments resulted in increases in catabolic processes, stimulation of mitochondrial and cytosolic metabolism, and decreases in chloroplast metabolism. Lower abundances of ribosomal proteins and translation factors under Low-N and Low-S corresponded with growth limitation. At the transcript level, the major and specific effect of Low-N was the enhancement of expression of defence and immunity genes. The main effect of chronic Low-S was a decrease in transcripts of genes involved in cell division, DNA replication, and cytoskeleton, and an increase in the expression of autophagy genes. This was consistent with a role of target-of-rapamycin kinase in the control of plant metabolism and cell growth and division under chronic Low-S. In addition, Low-S decreased the expression of several NLP transcription factors, which are master actors in nitrate sensing. Finally, both the transcriptome and proteome data indicated that Low-S repressed glucosinolate synthesis, and that Low-N exacerbated glucosinolate degradation. This showed the importance of glucosinolate as buffering molecules for N and S management. [ABSTRACT FROM AUTHOR]