Biology subjects

Helariutta, Y.

Publications and source records attributed to Helariutta, Y..

12 recordsLinked to original sources

bioRxiv2026

BpLAZY1A Mediates Transcriptional Polarity and Drives Adaxial Tension Wood Like Tissue Formation in Silver Birch (Betula pendula) Branches

A defining feature of most trees is the architectural distinction between a vertically growing main stem and laterally growing branches. This growth habit increases fitness by increasing photosynthetically active surface area and shading competitors. Here, we studied the weeping birch cultivar Betula pendula Youngii to uncover the mechanisms that maintain lateral branch growth. We identified a loss-of-function mutation in BpLAZY1A, a core component of the gravitropic signalling pathway, as the cause of the weeping phenotype. Forward genetic analysis demonstrated that the weeping phenotype is recessive in silver birch. Transgenic BpLAZY1A RNAi lines phenocopied Youngii, confirming the functional role of BpLAZY1A. Reporter analysis revealed that BpLAZY1A is expressed predominantly in gravity-sensing starch sheath cells, with occasional expression in the main stem phloem. Time lapse imaging revealed two distinct gravitropic responses during branch development - an early response associated with establishment of the branch apex gravitropic set-point angle, and a later response associated with polar reinforcement at the branch base. The main stem retained normal gravitropic responses in BpLAZY1A RNAi lines, indicating a branch-specific role for BpLAZY1A. Integrative analyses combining transcriptomics, chemical profiling, and histochemistry indicated that BpLAZY1A establishes adaxial-abaxial polarity during early branch development. In wild-type branches, pectin-rich tension wood-like tissue formed preferentially in the adaxial xylem, accompanied by adaxial expression of pectin- and tension wood-associated genes, including BpRRT1 and BpCOBRA-LIKE4. Auxin-related genes, including BpIAA29 and BpSAURs, were preferentially upregulated in the abaxial side. This transcriptional asymmetry was reduced in BpLAZY1A RNAi line 1, where tension wood-like tissue formed in both adaxial and abaxial sides of the xylem, indicating a loss of polarity. Together, these findings demonstrate that BpLAZY1A functions specifically in branch gravitropism and links spatially asymmetric gene expression with branch growth orientation and biomechanical reinforcement. Our results identify BpLAZY1A as a key regulator coordinating gravitropic signalling, tissue polarity, and the developmental biomechanics underlying lateral branch growth.

plant biology↗

bioRxiv2026

Rewiring vascular patterning through translational control in Arabidopsis

Plant vascular systems exhibit a wide developmental spectrum, from rigid woody tissues to soft, fleshy storage tissues. We show that increasing polyamine thermospermine transport into wild-type rootstocks, together with cytokinin, reprograms xylem identity from woody to fleshy in Arabidopsis. Our findings establish thermospermine as a mobile developmental signal and suggest a strategy for engineering plant vascular architecture.

plant biology↗

bioRxiv2023

Plasmodesmata act as unconventional membrane contact sites regulating inter-cellular molecular exchange in plants

Membrane contact sites (MCS) are fundamental for intracellular communication, but their role in intercellular communication remains unexplored. We show that in plants, plasmodesmata communication bridges function as atypical endoplasmic reticulum (ER)-plasma membrane (PM) tubular MCS, operating at cell-cell interfaces. Similar to other MCS, ER-PM apposition is controlled by a protein-lipid tethering complex, but uniquely, this serves intercellular communication. Combining high-resolution microscopy, molecular dynamics, pharmacological and genetic approaches, we show that cell-cell trafficking is modulated through the combined action of Multiple C2 domains and transmembrane domain proteins (MCTP) 3, 4, and 6 ER-PM tethers, and phosphatidylinositol-4-phosphate (PI4P) lipid. Graded PI4P amounts regulate MCTP docking to the PM, their plasmodesmata localization and cell-cell permeability. SAC7, an ER-localized PI4P-phosphatase, regulates MCTP4 accumulation at plasmodesmata and modulates cell-cell trafficking capacity in a cell-type specific manner. Our findings expand MCSs functions in information transmission, from intracellular to intercellular cellular activities. In briefPlant intercellular communication is regulated via tubular membrane contact through PI4P binding-ER-PM tether MCTP proteins HighlightsO_LIPlasmodesmata are unconventional ER/PM tubular contact sites located at cell-cell interface C_LIO_LIPlasmodesmata operate as control valves, modulating ER-PM contacts to regulate transport C_LIO_LIMCTP3, MCTP4, MCTP6 and PI4P tethering elements act as valve regulators C_LIO_LISAC7 PI4P phosphatase controls plasmodesmata MCS in a cell-type-specific manner C_LI

plant biology↗

bioRxiv2022

Pectin cell wall remodeling through PLL12 and callose deposition through polar CALS7 are necessary for long-distance phloem transport

In plants, the phloem distributes photosynthetic products for metabolism and storage over long distances. It relies on specialized cells, the sieve elements, which are enucleated and interconnected through large so-called sieve pores in their adjoining cell walls. Reverse genetics identified PECTATE LYASE LIKE 12 (PLL12) as critical for plant growth and development. Using genetic complementation, we establish that PLL12 enzyme activity is required exclusively late during cell differentiation. Physiological assays suggested a role specifically in phloem long distance transport while short distance symplastic transport does not require PLL12. Exploiting CALLOSE SYNTHASE 7 (CALS7) as a polar fluorescent marker, we identified structural defects in sieve pores in pll12. Due to the serial arrangement of sieve elements in the phloem, such defects should affect a plants ability specifically for symplastic transport over long distances.

plant biology↗

bioRxiv2021

In planta dynamics, transport-biases and endogenous functions of mobile siRNAs in Arabidopsis

O_LIIn RNA interference (RNAi), small-interfering (si)RNAs processed from double-stranded RNA guide ARGONAUTE(AGO) proteins to silence sequence-complementary RNA/DNA. Plant RNAi can propagate locally and systemically, but despite recent mechanistic advances, basic questions/hurdles remain unaddressed. For instance, RNAi is inferred to diffuse through plasmodesmata, yet how its dynamics in planta compares with that of established symplastic-diffusion markers remains unknown. Also unknown is why select siRNA species, or size-classes thereof, are recovered in RNAi-recipient tissues, yet only under some experimental settings. Finally, RNAi shootward movement in micro-grafted Arabidopsis - necessary to study its presumptive transgenerational effects- has not been achieved thus far and endogenous functions of mobile RNAi remain scarcely documented. C_LIO_LIFocusing on non-amplified RNAi in Arabidopsis, we show here that (i) transgenic RNAi-movement, although symplasmic, only partially recapitulates the diffusion pattern of free GFP in planta, (ii) the presence/absence of specific AGOs in incipient/traversed/recipient tissues likely explains the apparent siRNA-selectivity observed during vascular movement, (iii) stress application allows endo-siRNA translocation against the shoot-to-root phloem flow, and (iv) mobile endo-siRNAs generated from a single inverted-repeat(IR) locus, have the potential to regulate hundreds of transcripts. C_LIO_LIOur results close important knowledge-gaps, rationalize previously-noted inconsistencies between mobile RNAi settings, and provide a framework for functional endo-siRNA studies. C_LI

plant biology↗

bioRxiv2021

An Arabidopsis root phloem pole cell atlas reveals PINEAPPLE genes as transitioners to autotrophy

Single cell sequencing has recently allowed the generation of exhaustive root cell atlases. However, some cell types are elusive and remain underrepresented. Here, we use a second- generation single cell approach, where we zoom in on the root transcriptome sorting with specific markers to profile the phloem poles at an unprecedented resolution. Our data highlight the similarities among the developmental trajectories and gene regulatory networks communal to protophloem sieve element (PSE) adjacent lineages in relation to PSE enucleation, a key event in phloem biology. As a signature for early PSE-adjacent lineages, we have identified a set of DNA-binding with one finger (DOF) transcription factors, the PINEAPPLEs (PAPL), that act downstream of PHLOEM EARLY DOF (PEAR) genes, and are important to guarantee a proper root nutrition in the transition to autotrophy. Our data provide a holistic view of the phloem poles that act as a functional unit in root development.

plant biology↗

bioRxiv2021

Analysis of phloem trajectory links tissue maturation to cell specialization.

The mechanisms that allow cells in the plant meristem to coordinate tissue-wide maturation gradients with specialized cell networks are critical for indeterminate growth. Here, we reconstructed the protophloem developmental trajectory of 19 cells from cell birth to terminal differentiation at single cell resolution in the Arabidopsis root. We found that cellular specification is mediated near the stem cell niche by PHLOEM EARLY DOF (PEAR) transcription factors. However, the PEAR dependent differentiation program is repressed by a broad gradient of PLETHORA (PLT) transcription factors, which directly inhibit PEARs own direct target ALTERED PHLOEM DEVELOPMENT (APL). The dissipation of PLT gradient around 7 cells from the stem cell activates APL expression, and a subsequent transitional network that results in a "seesaw" pattern of mutual inhibition over developmental time. Together, we provide a mechanistic understanding of how morphogen-like maturation gradients interface with cell-type specific transcriptional regulators to stage cellular differentiation.

plant biology↗

bioRxiv2020

Blocking plasmodesmata in specific phloem cell types reduces axillary bud growth in Arabidopsis thaliana

The plasticity of above ground plant architecture depends on the regulated re-activation and growth of axillary meristems laid down in the axils of leaves along the stem, which often arrest as dormant buds. Plasmodesmata connecting plant cells might control the movement of regulators involved in this developmental switch. Constructs capable of occluding these structures were employed in phloem cell types, because of the importance of phloem in local and systemic trafficking. We show that over-accumulation of callose within companion cells of the Arabidopsis inflorescence reduces the growth rates of activated buds, but does not affect bud activation. Growth rate reductions were not dependent on the phloem-mobile strigolactone receptor, which regulates bud activation. Furthermore, there was no correlation with early bud sugar profiles, which can also affect bud activity and depend on phloem-mediated delivery. It is therefore possible that an as yet unknown mobile signal is involved in modulating branch growth rate.

plant biology↗

bioRxiv2020

Computational tools for serial block EM reveal differences in plasmodesmata distributions and wall environments

Plasmodesmata are small channels that connect plant cells. While recent technological advances have facilitated the analysis of the ultrastructure of these channels, there are limitations to efficiently addressing their presence over an entire cellular interface. Here, we highlight the value of serial block electron microscopy for this purpose. We developed a computational pipeline to study plasmodesmata distributions and we detect presence/absence of plasmodesmata clusters, pit fields, at the phloem unloading interfaces of Arabidopsis thaliana roots. Pit fields can be visualised and quantified. As the wall environment of plasmodesmata is highly specialised we also designed a tool to extract the thickness of the extracellular matrix at and outside plasmodesmata positions. We show and quantify clear wall thinning around plasmodesmata with differences between genotypes, namely in the recently published plm-2 sphingolipid mutant. Our tools open new avenues for quantitative approaches in the analysis of symplastic trafficking. Sentence summaryWe developed computational tools for serial block electron microscopy datasets to extract information on the spatial distribution of plasmodesmata over an entire cellular interface and on the wall environment the plasmodesmata are in.

plant biology↗

bioRxiv2020

GSK3 activity is a cell fate switch that balances the ratio of vascular cell type

The phloem transports photosynthetic assimilates and signalling molecules. It mainly consists of sieve elements (SEs), which act as "highways" for transport, and companion cells (CCs), which serve as "gates" to load/unload cargos. Though SEs and CCs function together, it remains unknown what determines the ratio of SE/CC in the phloem. In this study, we develop a novel culture system for CC differentiation named VISUAL-CC, which reconstitutes the SE-CC complex formation. Comparative expression analysis in VISUAL-CC reveals that SE and CC differentiation tends to show negative correlation, while total phloem differentiation is unchanged. This varying SE/CC ratio is largely dependent on GSK3 kinase activity. Indeed, gsk3 hextuple mutants possess much more SEs and less CCs in planta. Conversely, gsk3 gain-of-function mutants induced by phloem-specific promoter partially increased the CC ratio. Taken together, GSK3 activity appears to function as a cell fate switch in the phloem, thereby balancing the SE/CC ratio.

plant biology↗

bioRxiv2019

Structural imaging of native cryo-preserved secondary cell walls reveals presence of macrofibrils composed of cellulose, lignin and xylan.

The woody secondary cell walls of plants are the largest repository of renewable carbon biopolymers on the planet. These walls are made principally from cellulose and hemicelluloses and are impregnated with lignin. Despite their importance as the main load bearing structure for plant growth, as well as their industrial importance as both a material and energy source, the precise arrangement of these constituents within the cell wall is not yet fully understood. We have adapted low temperature scanning electron microscopy (cryo-SEM) for imaging the nanoscale architecture of angiosperm and gymnosperm cell walls in their native hydrated state. Our work confirms that cell wall macrofibrils, cylindrical structures with a diameter exceeding 10 nm, are a common feature of the native hardwood and softwood samples. We have observed these same structures in Arabidopsis thaliana secondary cell walls, enabling macrofibrils to be compared between mutant lines that are perturbed in cellulose, hemicellulose and lignin formation. Our analysis indicates that the macrofibrils in Arabidopsis cell walls are composed, at least partially, of cellulose, xylan and lignin. This study is a useful additional approach for investigating the native nanoscale architecture and composition of hardwood and softwood secondary cell walls and demonstrates the applicability of Arabidopsis genetic resources to relate fibril structure with wall composition and biosynthesis.

plant biology↗

bioRxiv2019

ELIMAKI locus is required for mechanosensing and proprioception in birch trees

The remarkable vertical and radial growth observed in tree species, encompasses a major physical challenge for wood forming tissues. To compensate with increasing size and weight, cambium-derived radial growth increases the stem width, thereby supporting the aerial body of trees. This feedback appears to be part of a so-called \"proprioception\" (1, 2) mechanism that controls plant size and biomass allocation. Yet, how trees experience or respond to mechanical stress derived from their own vertical loading, remains unknown. Here, we combined two strategies to dissect the proprioceptive response in birch. First, we show that in response to physical loading, trees promote radial growth with different magnitudes along the stem. Next, we identified a mutant cultivar (B. pubescens cv. Elimaki) in which the main stem shows normal vertical development, but collapses after three months. By inducing precocious flowering, we generated a backcrossed population (BC1) by producing two generations in 4 years. In his scheme, we uncovered a recessive trait (eki) that segregates and genetically maps with a Mendelian monogenic pattern. Unlike WT, eki is resistant to vertical mechanical stimulation. However, eki responds normally to the gravitropic stimulus by making tension wood. Before the collapse, cell size in eki is compromised resulting in radial growth defects, depending on stem height. Cell walls of developing xylem and phloem tissues have delayed differentiation in eki, and its tissues are softer compared to WT as indicated by atomic force microscopy (AFM). The transcriptomic profile of eki highlighted the overlap with that of the Arabidopsis response to touch. Taken together, our results suggest that the mechanical environment and cell wall properties of developing woody tissues, can significantly affect the growth responses to vertical loading thereby compromising their proprioceptive capacity. Additionally, we introduce a fast forward genetics strategy to dissect complex phenotypes in trees.

plant biology↗