Multiplexed in situ hybridization reveals distinct lineage identities for major and minor vein initiation during maize leaf development.

Leaves of flowering plants are characterized by diverse venation patterns. Patterning begins with the selection of vein-forming procambial initial cells from within the ground meristem of a developing leaf, a process which is considered to be auxin-dependent, and continues until veins are anatomically differentiated with functional xylem and phloem. At present, the mechanisms responsible for leaf venation patterning are primarily characterized in the model eudicot Arabidopsis thaliana which displays a reticulate venation network. However, evidence suggests that vein development may proceed via a different mechanism in monocot leaves where venation patterning is parallel. Here, we employed Molecular Cartography, a multiplexed in situ hybridization technique, to analyze the spatiotemporal localization of a subset of auxin-related genes and candidate regulators of vein patterning in maize leaves. We show how different combinations of auxin influx and efflux transporters are recruited during leaf and vein specification and how major and minor vein ranks develop with distinct identities. The localization of the procambial marker PIN1a and the spatial arrangement of procambial initial cells that give rise to major and minor vein ranks further suggests that vein spacing is prepatterned across the medio-lateral leaf axis prior to accumulation of the PIN1a auxin transporter. In contrast, patterning in the adaxial-abaxial axis occurs progressively, with markers of xylem and phloem gradually becoming polarized as differentiation proceeds. Collectively, our data suggest that both lineage- and position-based mechanisms may underpin vein patterning in maize leaves.

The WIP6 transcription factor TOO MANY LATERALS specifies vein type in C4 and C3 grass leaves.

Grass leaves are invariantly strap shaped with an elongated distal blade and a proximal sheath that wraps around the stem. Underpinning this shape is a scaffold of leaf veins, most of which extend in parallel along the proximo-distal leaf axis. Differences between species are apparent both in the vein types that develop and in the distance between veins across the medio-lateral leaf axis. A prominent engineering goal is to increase vein density in leaves of C3 photosynthesizing species to facilitate the introduction of the more efficient C4 pathway. Here, we discover that the WIP6 transcription factor TOO MANY LATERALS (TML) specifies vein rank in both maize (C4) and rice (C3). Loss-of-function tml mutations cause large lateral veins to develop in positions normally occupied by smaller intermediate veins, and TML transcript localization in wild-type leaves is consistent with a role in suppressing lateral vein development in procambial cells that form intermediate veins. Attempts to manipulate TML function in rice were unsuccessful because transgene expression was silenced, suggesting that precise TML expression is essential for shoot viability. This finding may reflect the need to prevent the inappropriate activation of downstream targets or, given that transcriptome analysis revealed altered cytokinin and auxin signaling profiles in maize tml mutants, the need to prevent local or general hormonal imbalances. Importantly, rice tml mutants display an increased occupancy of veins in the leaf, providing a step toward an anatomical chassis for C4 engineering. Collectively, a conserved mechanism of vein rank specification in grass leaves has been revealed.

The pandemic seen through the eyes of the youngest people: evaluating psychological impact of the early COVID-19 related confinement on children and adolescents through the analysis of drawings and of an e-survey on their parents.

BACKGROUND: COVID-19 is having a significant impact on long term children' and adolescents' psychological health. We aimed to evaluate the direct early psychological and behavioural signs related to the COVID-19 pandemic outbreak and related confinement on children and adolescents. METHODS: Children and adolescents' drawings were collected for a limited time window (16th March-10th April 2020) and analyzed. Their parents were asked in the following month to answer a qualitative e-survey on somatic complaints and behavioral changes of the participating children/adolescents. RESULTS: Ninety-eight drawings by children/adolescents (mean age 7.01±2.83 years) were analysed. Analyses of the 98 drawings reported signs of trauma in all (of them, 60.2% with moderate-to high levels). Children aged 3-5 years were more impacted, followed by preadolescents/adolescents aged 11-17 years. Parents reported somatic complaints in the 71.1% of their children/adolescents: the most frequent were increased appetite (35.6%), abdominal pain (20.0%) and headache (20.0%). Behavioral changes were observed in 77.8% of subjects: increased appetite (35.6%), abdominal pain (20.0%) and headache (20.0%) were more represented. CONCLUSIONS: Early psychological distress related to COVID-19 pandemic was observed both in children and in adolescents by the analysis of drawings and confirmed by their parents. Implementation of mental health-care services for preventing future psychopathological problems is mandatory.

Developmental regulation of leaf venation patterns: monocot versus eudicots and the role of auxin.

Organisation and patterning of the vascular network in land plants varies in different taxonomic, developmental and environmental contexts. In leaves, the degree of vascular strand connectivity influences both light and CO2 harvesting capabilities as well as hydraulic capacity. As such, developmental mechanisms that regulate leaf venation patterning have a direct impact on physiological performance. Development of the leaf venation network requires the specification of procambial cells within the ground meristem of the primordium and subsequent proliferation and differentiation of the procambial lineage to form vascular strands. An understanding of how diverse venation patterns are manifest therefore requires mechanistic insight into how procambium is dynamically specified in a growing leaf. A role for auxin in this process was identified many years ago, but questions remain. In this review we first provide an overview of the diverse venation patterns that exist in land plants, providing an evolutionary perspective. We then focus on the developmental regulation of leaf venation patterns in angiosperms, comparing patterning in eudicots and monocots, and the role of auxin in each case. Although common themes emerge, we conclude that the developmental mechanisms elucidated in eudicots are unlikely to fully explain how parallel venation patterns in monocot leaves are elaborated.

Arabidopsis thaliana myosin XIK is recruited to the Golgi through interaction with a MyoB receptor.

Plant cell organelles are highly mobile and their positioning play key roles in plant growth, development and responses to changing environmental conditions. Movement is acto-myosin dependent. Despite controlling the dynamics of several organelles, myosin and myosin receptors identified so far in Arabidopsis thaliana generally do not localise to the organelles whose movement they control, raising the issue of how specificity is determined. Here we show that a MyoB myosin receptor, MRF7, specifically localises to the Golgi membrane and affects its movement. Myosin XI-K was identified as a putative MRF7 interactor through mass spectrometry analysis. Co-expression of MRF7 and XI-K tail triggers the relocation of XI-K to the Golgi, linking a MyoB/myosin complex to a specific organelle in Arabidopsis. FRET-FLIM confirmed the in vivo interaction between MRF7 and XI-K tail on the Golgi and in the cytosol, suggesting that myosin/myosin receptor complexes perhaps cycle on and off organelle membranes. This work supports a traditional mechanism for organelle movement where myosins bind to receptors and adaptors on the organelle membranes, allowing them to actively move on the actin cytoskeleton, rather than passively in the recently proposed cytoplasmic streaming model.

Multiple object-tracking isolates feedback-specific load in attention and learning.

Feedback is beneficial for learning. Nevertheless, it remains unclear whether (i) feedback draws attentional resources when integrated and (ii) the benefits of feedback for learning can be demonstrated using an attention-based task. We therefore (i) isolated feedback-specific load from task-specific load via individual differences in attention resource capacity and (ii) examined the effect of trial-by-trial feedback (i.e., present vs. absent) on learning a multiple object-tracking (MOT) paradigm. We chose MOT because it is a robust measure of attention resource capacity. In Study 1 participants tracked one (i.e., lowest attentional load condition) through four target items (i.e., highest load condition) among eight total items. One group (n = 32) received trial-by-trial feedback whereas the other group (n = 32) did not. The absence of feedback resulted in better MOT performance compared with the presence of feedback. Moreover, the difference in MOT capability between groups increased as the task-specific attentional load increased. These findings suggest that feedback integration requires attentional resources. Study 2 examined whether the absence (n = 19) or presence (n = 19) of feedback affects learning on the same MOT task across four testing days. When holding task-specific load constant, improvement in MOT was greater with feedback than without. Although this study is the first to isolate feedback-specific load in attention with MOT, more evidence is needed to demonstrate how the benefits of feedback translate to improvement on an attention-based task. These findings encourage future research to further explore the interaction between feedback, attention and learning.

Plant organelle dynamics: cytoskeletal control and membrane contact sites.

Contents Summary 381 I. Introduction 381 II. Basic movement characteristics 382 III. Actin and associated motors, myosins, play a primary role in plant organelle movement and positioning 382 IV. Mechanisms of myosin recruitment: a tightly regulated system? 384 V. Microtubules, associated motors and interplay with actin 386 VI. Role of organelle interactions: tales of tethers 387 VII. Summary model to describe organelle movement in higher plants 390 VIII. Why is organelle movement important? 390 IX. Conclusions and future perspectives 391 Acknowledgements 391 References 391 SUMMARY: Organelle movement and positioning are correlated with plant growth and development. Movement characteristics are seemingly erratic yet respond to external stimuli including pathogens and light. Given these clear correlations, we still do not understand the specific roles that movement plays in these processes. There are few exceptions including organelle inheritance during cell division and photorelocation of chloroplasts to prevent photodamage. The molecular and biophysical components that drive movement can be broken down into cytoskeletal components, motor proteins and tethers, which allow organelles to physically interact with one another. Our understanding of these components and concepts has exploded over the past decade, with recent technological advances allowing an even more in-depth profiling. Here, we provide an overview of the cytoskeletal and tethering components and discuss the mechanisms behind organelle movement in higher plants.

Plant ER geometry and dynamics: biophysical and cytoskeletal control during growth and biotic response.

The endoplasmic reticulum (ER) is an intricate and dynamic network of membrane tubules and cisternae. In plant cells, the ER 'web' pervades the cortex and endoplasm and is continuous with adjacent cells as it passes through plasmodesmata. It is therefore the largest membranous organelle in plant cells. It performs essential functions including protein and lipid synthesis, and its morphology and movement are linked to cellular function. An emerging trend is that organelles can no longer be seen as discrete membrane-bound compartments, since they can physically interact and 'communicate' with one another. The ER may form a connecting central role in this process. This review tackles our current understanding and quantification of ER dynamics and how these change under a variety of biotic and developmental cues.