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Efroni, I.

Publications and source records attributed to Efroni, I..

11 recordsLinked to original sources

Wounding-Induced Redirection of Sugar Transport Fuels Tissue Repair

Wounding triggers growth programs to restore damaged tissues, creating a local surge in metabolic demand. How resource recruitment is modified to meet this demand is unclear. Here, we show that regeneration of dissected root tips is dependent on photosynthetic sucrose in a dose-dependent manner, although sucrose itself is excluded from the injury site. High-resolution tracking of Glifon, a novel live glucose reporter, reveals that glucose accumulates near the cut. Glucose accumulation required the apoplasmic sugar transport components CELL WALL INVERTASE (CWINV) and SUGAR TRANSPORTER PROTEINS (STP), which were rapidly induced by wounding. Loss of CWINV or STP function compromised root repair, particularly under limited sucrose availability, whereas increased STP13 gene dosage enhanced repair rates. Similar sugar transport genes were activated in other wounding contexts and promoted wound-induced adventitious root initiation. We propose that injury elicits a proactive local shift in sugar flow to promote resource recruitment and sustain tissue repair. Significance StatementPlants grow under constant physical assault and have evolved mechanisms to repair damaged organs. Wound repair increases demand for sugars to fuel cell division and growth, but unlike animals, plants cannot dilate blood vessels and rely on fixed phloem tubes. Our work shows how plants solve this problem. We uncover a circuit that blocks entry of the transported sugar sucrose into the wound site, converting it to glucose in the cell wall space, which is then rapidly imported into injury-adjacent cells. This wound-induced mechanism precedes repair, creating a proactive "sugar sink" that draws carbohydrates toward the damaged zone. Boosting this mechanism increases wound repair rates, suggesting this mechanism allows plants to control resource distribution, balancing growth and wound repair.

plant biology↗

A deep-time landscape of plant cis-regulatory sequence evolution

Developmental gene function is often conserved over deep time, but cis-regulatory sequence conservation is difficult to identify. Rapid sequence turnover, paleopolyploidy, structural variation, and limited phylogenomic sampling have impeded conserved non-coding sequence (CNS) discovery. Using Conservatory, an algorithm that leverages microsynteny and iterative alignments to map CNS-gene associations over evolution, we uncovered [~]2.3 million CNSs, including over 3,000 predating angiosperms, from 284 plant species spanning 300 million years of diversification. Ancient CNSs were enriched near developmental regulators, and mutating CNSs near HOMEOBOX genes produced strong phenotypes. Tracing CNS evolution uncovered key principles: CNS spacing varies, but order is conserved; genomic rearrangements form new CNS-gene associations; and ancient CNSs are preferentially retained among paralogs, but are often lost as cohorts or evolve into lineage-specific CNSs. One Sentence SummaryConservatory maps ancient cis-regulatory elements and uncovers regulatory evolution dynamics.

plant biology↗

TOTEM: A web TOol for Tissue-EnrichMent analysis on gene lists

Analysis of spatiotemporal patterns of gene expression is crucial to decode biological systems responses. High-throughput sequencing allows in-depth transcriptome analyses and experimental designs, providing valuable reference expression atlases. Specifically, testing overrepresentations of tissue-specific genes based on these atlases can provide valuable insights; however, such an approach is not accessible to inexperienced users. Here, we introduce TOTEM (TOol for Tissue-EnrichMent), a web tool designed to calculate enrichment values per tissue by identifying tissue-specific genes from an organ/organism of interest given a user gene list. Results are visually represented, and users gene classified. The utility of TOTEM is manifest when using integrated single cell expression atlases, enabling the study of complicated tissues, with the maximum possible resolution. Its effectiveness is validated by the study of BRL3 role in stress specifically from the vascular tissues. Finally, TOTEMs modular design allows for continual integration of new experiments. TOTEM can be freely accessed at: https://totemwebtool.com.

plant biology↗

Restriction of intercellular communication is required for organ regeneration

The excision of the root tip, including the stem cell niche, triggers rapid regeneration from remnant cells in the stump. In plants, cell identity relies on positional information transported through cytoplasmatic bridges known as plasmodesmata. However, how such information is reset to allow the regeneration of lost identities is unknown. Here, we show that the movement of mobile signals is temporarily restricted near the incision site and that regeneration-induced members of the LATERAL ORGAN DOMAIN (LBD) plant-specific developmental regulators are necessary and sufficient for this restriction. Regeneration was disrupted in high-order lbd mutants but was restored by transient localized plasmodesmata closure. We propose that LBD-mediated modulation of intercellular connectivity is crucial for plant regeneration and may have widespread roles in de novo pattern formation. One Sentence SummaryPlant-specific DNA binding genes mediate a transient restriction of intercellular communication to allow root regeneration

plant biology↗

Mapping of the Classical Mutation rosette Highlights a Role for Calcium in Wound-induced Rooting

Removal of the root system induces the formation of new roots from the remaining shoot. This process is primarily controlled by the phytohormone auxin which interacts with other signals in a yet unresolved manner. Here, we study the classical tomato mutation rosette (ro) which lacks shoot-borne roots. ro plants were severely inhibited in the formation of wound-induced roots and have reduced rates of auxin transport. We mapped ro to the tomato ortholog of the Arabidopsis thaliana BIG and the mammalians UBR4/p600. RO/BIG is a large protein of unknown biochemical function. In A. thaliana, BIG was implicated in the regulation of auxin transport and calcium homeostasis. We show that exogenous calcium inhibits wound-induced root formation in both tomato and A. thaliana ro/big mutants. Exogenous calcium antagonized the root-promoting effects of the auxin IAA, but not of 2,4-D, an auxin analog that is not recognized by the polar transport machinery, and accumulation of the auxin transporter PIN1 was sensitive to calcium levels in the ro/big mutants. Consistent with a role for calcium in mediating auxin transport, both ro/big mutants and calcium-treated wild-type plants were hypersensitive to treatment with polar auxin transport inhibitors. Subcellular localization of BIG suggests that like its mammalian ortholog, it is associated with the endoplasmic reticulum (ER). Analysis of subcellular morphology revealed that ro/big mutants exhibited disruption in cytoplasmic streaming. We suggest that RO/BIG maintain auxin flow by stabilizing PIN membrane localization, possibly by attenuating the inhibitory effect of Ca2+ on cytoplasmic streaming.

plant biology↗

CLASS-II KNOX genes coordinate spatial and temporal patterns of the tomato ripening

Ripening is a complex developmental change of a mature organ, the fruit. In plants like a tomato, it involves softening, pigmentation, and biosynthesis of metabolites beneficial for the human diet. Examination of the transcriptional changes towards ripening suggests that redundant uncharacterized factors may be involved in the coordination of the ripening switch. Previous studies have demonstrated that Arabidopsis CLASS-II KNOX genes play a significant role in controlling the maturation of siliques and their transition to senescence. Here we examined the combined role of all four tomato CLASS-II KNOX genes in the maturation and ripening of fleshy fruits using an artificial microRNA targeting them simultaneously. As expected, the knockdown plants (35S::amiR-TKN-CL-II) exhibited leaves with increased complexity, reminiscent of the leaf phenotype of plants overexpressing CLASS-I KNOX, which antagonize CLASS-II KNOX gene functions. The fruits of 35S::amiR-TKN-CL-II plants were notably smaller than the control. While their internal gel/placenta tissue softened and accumulated the typical pigmentation, the pericarp color break took place ten days later than control, and eventually, it turned yellow instead of red. Additionally, the pericarp of 35S::amiR-TKN-CL-II fruits remained significantly firmer than control even after three weeks of shelf storage. Strikingly, the 35S::amiR-TKN-CL-II fruits showed early ethylene release and respiration peak, but these were correlated only with liquefaction and pigmentation of the internal tissues. Our findings suggest that CLASS-II KNOX genes are required to coordinate the spatial and temporal patterns of tomato fruit ripening. One sentence summaryTomato CLASS-II KNOX genes play antagonistic roles in the regulation of ripening at the internal fruit domains and pericarp.

developmental biology↗

Auxin requirements for a meristematic state in roots depend on a dual brassinosteroid function

The organization of the root meristem is maintained by a complex interplay between plant hormones signaling pathways that both interpret and determine their accumulation and distribution. Brassinosteroids (BR) and auxin signaling pathways control the number of meristematic cells in the Arabidopsis root, via an interaction that appears to involve contradicting molecular outcomes, with BR promoting auxin signaling input but also repressing its output. However, whether this seemingly incoherent effect is significant for meristem function is unclear. Here, we established that a dual effect of BR on auxin, with BR simultaneously promoting auxin biosynthesis and repressing auxin transcriptional output, is essential for meristem maintenance. Blocking BR-induced auxin synthesis resulted in rapid BR-mediated meristem loss. Conversely, plants with reduced BR levels were resistant to loss of auxin biosynthesis and these meristems maintained their normal morphology despite a 10-fold decrease in auxin levels. In agreement, injured root meristems which rely solely on local auxin synthesis, regenerated when both auxin and BR synthesis were inhibited. Use of BIN2 as a tool to selectively inhibit BR signaling, revealed meristems with distinct phenotypes depending on the perturbed tissue; meristem reminiscent of BR-deficient mutants or of high BR exposure. This enabled mapping BR-auxin interactions to the outer epidermis and lateral root cap tissues, and demonstrated the essentiality of BR signaling in these tissues for meristem maintenance. BR activity in internal tissues however, proved necessary to control BR homeostasis. Together, we demonstrate a basis for inter-tissue coordination and how a critical ratio between these hormones determines the meristematic state.

plant biology↗

A conserved superlocus regulates above- and belowground root initiation

During plant post-embryonic growth new meristems and associated stem cells form in different development contexts in order to respond to environmental cues. While underground lateral roots initiate from designated cells in the main root, an unknown mechanism allows cells to bypass the root/shoot identity trajectory and generate shoot-borne-roots. Using single-cell profiling of tomato (Solanum lycoperiscum) stems we isolated a rare transient cell population that serve as progenitors for shoot-borne-root meristems. Analysis of this population identified a transcription factor required for the formation of shoot-borne-roots which we named SHOOT BORNE ROOTLESS (SBRL). Evolutionary analysis revealed that SBRL function is deeply conserved in angiosperms and that it arose as part of an ancient duplicated superlocus, only lost in root-less plants, containing both shoot-borne and lateral root initiation regulators. We propose that the ability to activate a common transition state with context-specific regulators allows the remarkable developmental plasticity found in plants. One Sentence SummaryHighly conserved superlocus of LBD genes, acting within an early transition identity, regulates shoot-borne and lateral root formation.

plant biology↗

Collaboration of multiple pathways in making a compound leaf

O_LIThe variability in leaf form in nature is immense. Leaf patterning occurs by differential growth that occurs during a limited window of morphogenetic activity at the leaf marginal meristem. While many regulators have been implicated in the designation of the morphogenetic window and in leaf patterning, how these effectors interact to generate a particular form is still not well understood. C_LIO_LIWe addressed the interaction among different effectors of tomato compound leaf development, using genetic and molecular analyses. C_LIO_LIMutations in the tomato auxin response factor SlARF5/SlMP, which promotes leaflet formation, suppressed the increased leaf complexity of mutants with extended morphogenetic window. Impaired activity of the NAC/CUC transcription factor GOBLET (GOB), which specifies leaflet boundaries, also reduced leaf complexity in these backgrounds. Analysis of genetic interactions showed that the patterning factors SlMP, GOB and the MYB transcription factor LYRATE (LYR) act in parallel to promote leaflet formation. C_LIO_LIThis work places an array of developmental regulators in a morphogenetic context. It reveals how organ-level differentiation rate and local growth are coordinated to sculpture an organ. These concepts and findings are applicable to other plant species and developmental processes that are regulated by patterning and differentiation. C_LI

plant biology↗

A dynamic pattern of local auxin sources is required for root regeneration

Following removal of its stem cell niche, the root meristem can regenerate by recruitment of remnant cells from the stump. Regeneration is initiated by rapid accumulation of auxin near the injury site but the source of this auxin is unknown. Here, we show that auxin accumulation arises from the activity of multiple auxin biosynthetic sources that are newly specified near the cut site and that their continuous activity is required for the regeneration process. Auxin synthesis is highly localized and PIN-mediate transport is dispensable for auxin accumulation and tip regeneration. Roots lacking the activity of the regeneration competence factor ERF115, or that are dissected at a zone of low-regeneration potential, fail to activate local auxin sources. Remarkably, restoring auxin supply is sufficient to confer regeneration capacity to these recalcitrant tissues. We suggest that regeneration competence relies on the ability to specify new local auxin sources in a precise spatio-temporal pattern.

plant biology↗

Deep conservation of response element variants regulating plant hormonal responses

Phytohormones regulate many aspects of plant life by activating transcription factors (TF) that bind sequence-specific response elements (RE) in regulatory regions of target genes. Despite their short length, REs are degenerate with a core of just 3-4bps. This degeneracy is paradoxical, as it reduces specificity, and REs are extremely common in the genome. To study whether RE degeneracy might serve a biological function we developed an algorithm for the detection of regulatory sequence conservation, and applied it to phytohormones REs in 45 angiosperms. Surprisingly, we found that specific RE variants are highly conserved in core hormone response genes. Experimental evidence showed that specific variants act to regulate the magnitude and spatial profile of hormonal response in Arabidopsis and tomato. Our results suggest that hormone-regulated TFs bind a spectrum of REs, each coding for a distinct transcriptional response profile. Our approach is applicable for precise editing and rational promoter design.

plant biology↗