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1.
Curr Biol ; 33(5): 926-939.e9, 2023 03 13.
Artigo em Inglês | MEDLINE | ID: mdl-36805125

RESUMO

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 complementations, we established that PLL12 is required exclusively late during sieve element differentiation. Structural homology modeling, enzyme inactivation, and overexpression suggest a vital role for PLL12 in sieve-element-specific pectin remodeling. While short distance symplastic diffusion is unaffected, the pll12 mutant is unable to accommodate sustained plant development due to an incapacity to accommodate increasing hydraulic demands on phloem long-distance transport as the plant grows-a defect that is aggravated when combined with another sieve-element-specific mutant callose synthase 7 (cals7). Establishing CALS7 as a specific sieve pore marker, we investigated the subcellular dynamics of callose deposition in the developing sieve plate. Using fluorescent CALS7 then allowed identifying structural defects in pll12 sieve pores that are moderate at the cellular level but become physiologically relevant due to the serial arrangement of sieve elements in the sieve tube. Overall, pectin degradation through PLL12 appears subtle in quantitative terms. We therefore speculate that PLL12 may act as a regulator to locally remove homogalacturonan, thus potentially enabling further extracellular enzymes to access and modify the cell wall during sieve pore maturation.


Assuntos
Arabidopsis , Arabidopsis/metabolismo , Floema/metabolismo , Glucanos/metabolismo , Plantas/metabolismo
2.
Nat Plants ; 8(8): 954-970, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35927456

RESUMO

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 common 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.


Assuntos
Arabidopsis , Floema , Arabidopsis/genética , Diferenciação Celular , Redes Reguladoras de Genes , Floema/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
3.
Curr Biol ; 32(9): 1883-1894.e7, 2022 05 09.
Artigo em Inglês | MEDLINE | ID: mdl-35320706

RESUMO

Wound healing is a fundamental property of plants and animals that requires recognition of cellular damage to initiate regeneration. In plants, wounding activates a defense response via the production of jasmonic acid and a regeneration response via the hormone auxin and several ethylene response factor (ERF) and NAC domain-containing protein (ANAC) transcription factors. To better understand how plants recognize damage and initiate healing, we searched for factors upregulated during the horticulturally relevant process of plant grafting and found four related DNA binding with one finger (DOF) transcription factors, HIGH CAMBIAL ACTIVITY2 (HCA2), TARGET OF MONOPTEROS6 (TMO6), DOF2.1, and DOF6, whose expression rapidly activated at the Arabidopsis graft junction. Grafting or wounding a quadruple hca2, tmo6, dof2.1, dof6 mutant inhibited vascular and cell-wall-related gene expression. Furthermore, the quadruple dof mutant reduced callus formation, tissue attachment, vascular regeneration, and pectin methylesterification in response to wounding. We also found that activation of DOF gene expression after wounding required auxin, but hormone treatment alone was insufficient for their induction. However, modifying cell walls by enzymatic digestion of cellulose or pectin greatly enhanced TMO6 and HCA2 expression, whereas genetic modifications to the pectin or cellulose matrix using the PECTIN METHYLESTERASE INHIBITOR5 overexpression line or korrigan1 mutant altered TMO6 and HCA2 expression. Changes to the cellulose or pectin matrix were also sufficient to activate the wound-associated ERF115 and ANAC096 transcription factors, suggesting that cell-wall damage represents a common mechanism for wound perception and the promotion of tissue regeneration.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Parede Celular/metabolismo , Celulose , Regulação da Expressão Gênica de Plantas , Hormônios/metabolismo , Ácidos Indolacéticos/metabolismo , Pectinas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Cicatrização
4.
Science ; 374(6575): eaba5531, 2021 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-34941412

RESUMO

In the plant meristem, tissue-wide maturation gradients are coordinated with specialized cell networks to establish various developmental phases required for indeterminate growth. Here, we used single-cell transcriptomics to reconstruct the protophloem developmental trajectory from the birth of cell progenitors to terminal differentiation in the Arabidopsis thaliana root. PHLOEM EARLY DNA-BINDING-WITH-ONE-FINGER (PEAR) transcription factors mediate lineage bifurcation by activating guanosine triphosphatase signaling and prime a transcriptional differentiation program. This program is initially repressed by a meristem-wide gradient of PLETHORA transcription factors. Only the dissipation of PLETHORA gradient permits activation of the differentiation program that involves mutual inhibition of early versus late meristem regulators. Thus, for phloem development, broad maturation gradients interface with cell-type-specific transcriptional regulators to stage cellular differentiation.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/citologia , Floema/citologia , Floema/crescimento & desenvolvimento , Raízes de Plantas/citologia , Fatores de Transcrição/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Diferenciação Celular , Proteínas de Ligação ao GTP/genética , Proteínas de Ligação ao GTP/metabolismo , Meristema/citologia , Floema/genética , Floema/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , RNA-Seq , Transdução de Sinais , Análise de Célula Única , Fatores de Transcrição/genética , Transcriptoma
5.
Annu Rev Plant Biol ; 72: 847-866, 2021 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-33730513

RESUMO

Single-cell approaches are quickly changing our view on biological systems by increasing the spatiotemporal resolution of our analyses to the level of the individual cell. The field of plant biology has fully embraced single-cell transcriptomics and is rapidly expanding the portfolio of available technologies and applications. In this review, we give an overview of the main advances in plant single-cell transcriptomics over the past few years and provide the reader with an accessible guideline covering all steps, from sample preparation to data analysis. We end by offering a glimpse of how these technologies will shape and accelerate plant-specific research in the near future.


Assuntos
Análise de Célula Única , Transcriptoma , Biologia Computacional , Plantas/genética , Análise de Sequência de RNA
6.
Curr Biol ; 29(3): 520-529.e6, 2019 02 04.
Artigo em Inglês | MEDLINE | ID: mdl-30686737

RESUMO

To create a three-dimensional structure, plants rely on oriented cell divisions and cell elongation. Oriented cell divisions are specifically important in procambium cells of the root to establish the different vascular cell types [1, 2]. These divisions are in part controlled by the auxin-controlled TARGET OF MONOPTEROS5 (TMO5) and LONESOME HIGHWAY (LHW) transcription factor complex [3-7]. Loss-of-function of tmo5 or lhw clade members results in strongly reduced vascular cell file numbers, whereas ectopic expression of both TMO5 and LHW can ubiquitously induce periclinal and radial cell divisions in all cell types of the root meristem. TMO5 and LHW interact only in young xylem cells, where they promote expression of two direct target genes involved in the final step of cytokinin (CK) biosynthesis, LONELY GUY3 (LOG3) and LOG4 [8, 9] Therefore, CK was hypothesized to act as a mobile signal from the xylem to trigger divisions in the neighboring procambium cells [3, 6]. To unravel how TMO5/LHW-dependent cytokinin regulates cell proliferation, we analyzed the transcriptional responses upon simultaneous induction of both transcription factors. Using inferred network analysis, we identified AT2G28510/DOF2.1 as a cytokinin-dependent downstream target gene. We further showed that DOF2.1 controls specific procambium cell divisions without inducing other cytokinin-dependent effects such as the inhibition of vascular differentiation. In summary, our results suggest that DOF2.1 and its closest homologs control vascular cell proliferation, thus leading to radial expansion of the root.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/fisiologia , Proliferação de Células/genética , Citocininas/metabolismo , Fatores Genéricos de Transcrição/genética , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Câmbio/fisiologia , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Transativadores/genética , Transativadores/metabolismo , Fatores Genéricos de Transcrição/metabolismo , Xilema/fisiologia
7.
Nature ; 565(7740): 490-494, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30626969

RESUMO

Apical growth in plants initiates upon seed germination, whereas radial growth is primed only during early ontogenesis in procambium cells and activated later by the vascular cambium1. Although it is not known how radial growth is organized and regulated in plants, this system resembles the developmental competence observed in some animal systems, in which pre-existing patterns of developmental potential are established early on2,3. Here we show that in Arabidopsis the initiation of radial growth occurs around early protophloem-sieve-element cell files of the root procambial tissue. In this domain, cytokinin signalling promotes the expression of a pair of mobile transcription factors-PHLOEM EARLY DOF 1 (PEAR1) and PHLOEM EARLY DOF 2 (PEAR2)-and their four homologues (DOF6, TMO6, OBP2 and HCA2), which we collectively name PEAR proteins. The PEAR proteins form a short-range concentration gradient that peaks at protophloem sieve elements, and activates gene expression that promotes radial growth. The expression and function of PEAR proteins are antagonized by the HD-ZIP III proteins, well-known polarity transcription factors4-the expression of which is concentrated in the more-internal domain of radially non-dividing procambial cells by the function of auxin, and mobile miR165 and miR166 microRNAs. The PEAR proteins locally promote transcription of their inhibitory HD-ZIP III genes, and thereby establish a negative-feedback loop that forms a robust boundary that demarks the zone of cell division. Taken together, our data establish that during root procambial development there exists a network in which a module that links PEAR and HD-ZIP III transcription factors integrates spatial information of the hormonal domains and miRNA gradients to provide adjacent zones of dividing and more-quiescent cells, which forms a foundation for further radial growth.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/citologia , Arabidopsis/genética , Câmbio/crescimento & desenvolvimento , Câmbio/genética , Regulação da Expressão Gênica de Plantas , Fatores de Transcrição/metabolismo , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/biossíntese , Proteínas de Arabidopsis/genética , Câmbio/citologia , Câmbio/metabolismo , Divisão Celular/genética , Sinais (Psicologia) , Citocininas/metabolismo , Ácidos Indolacéticos/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Floema/citologia , Floema/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Raízes de Plantas/citologia , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Transdução de Sinais , Fatores de Transcrição/biossíntese , Fatores de Transcrição/genética , Transcrição Gênica
8.
J Plant Res ; 131(1): 31-36, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29204753

RESUMO

Plant vasculature consists of two major conductive cell types, xylem tracheary elements and phloem sieve elements (SEs). Both cell types undergo a highly specialized differentiation process. The root meristem of Arabidopsis displays a stereotypical anatomy in which the central vasculature is surrounded by concentric layers of outer tissues. Each cell file is derived from stem cells located in the root tip. A series of formative and proliferative divisions take place in the meristem; these are followed by cell expansion and differentiation. Protophloem differentiation is unique in being complete only 20-25 cells away from the first stem cell, and during the differentiation process the cells lose several organelles, including the nucleus, while the remaining organelles are rearranged. Defects in SE development have been shown to result in impaired auxin transport and response and therefore systemically affect root growth. Although a few genes have been demonstrated to function in phloem development, detailed analyses and a comprehensive understanding of sieve element development (i.e. how often the stem cells divide, how frequently enucleation takes place, and how SE development is coordinated between cell division and differentiation on a molecular level) are still lacking. Advanced live-imaging techniques which enable prolonged time-lapse captures of root tip growth as well as single-cell transcriptomic analysis of the 20-25 cells in the SE file could help resolve these questions. In addition, understanding the interplay between the PLETHORA (PLT) gradient, which is known to govern the root zonation, and phloem development within the root meristem could shed light on the rapidity of SE differentiation and its importance to the meristem.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/fisiologia , Diferenciação Celular , Regulação da Expressão Gênica de Plantas , Meristema/fisiologia , Floema/fisiologia , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Modelos Genéticos
9.
Curr Opin Plant Biol ; 35: 23-29, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27794261

RESUMO

Two major conducting tissues in plants, phloem and xylem, are composed of highly specialized cell types adapted to long distance transport. Sieve elements (SEs) in the phloem display a thick cell wall, callose-rich sieve plates and low cytoplasmic density. SE differentiation is driven by selective autolysis combined with enucleation, after which the plasma membrane and some organelles are retained. By contrast, differentiation of xylem tracheary elements (TEs) involves complete clearance of the cellular components by programmed cell death followed by autolysis of the protoplast; this is accompanied by extensive deposition of lignin and cellulose in the cell wall. Emerging molecular data on TE and SE differentiation indicate a central role for NAC and MYB type transcription factors in both processes.


Assuntos
Diferenciação Celular , Floema/fisiologia , Desenvolvimento Vegetal/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Xilema/fisiologia , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo
10.
Plant Signal Behav ; 9(9): e29581, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25763696

RESUMO

Forisomes are specialized multimeric protein complexes found only in the papilionoid legumes. They undergo a reversible conformational change in response to phloem injury to enable the occlusion of sieve tubes, thus preventing the loss of photoassimilates. The individual subunits are designated by the letters SEO-F (sieve element occlusion by forisomes) and are part of the larger SEO protein family, which also includes the typical P-proteins found in most dicots and some monocots. When specific SEO-F subunits from different species are expressed in a heterologous background, they self-assemble into fully-functional artificial forisomes. However, with the exception of basal species such as Dipteryx panamensis, the geometry of these artificial forisomes differs from that of their native counterparts. Studies involving SEO-F proteins from the model legume Medicago truncatula have shown that a combination of 3 of the 4 subunits can fine-tune the geometry of artificial forisomes. However, MtSEO-F3 was excluded from these studies because it was not incorporated into either the native or artificial forisomes in our original experiments. In this addendum, we present further data concerning the interactive properties of the SEO-F proteins and confirm that all 4 MtSEO-F proteins interact in all possible pairwise combinations. These data indicate that the exclusion of MtSEO-F3 from the compact forisome may reflect the steric hindrance of binding sites rather than an inability to interact with other forisome subunits.


Assuntos
Medicago truncatula/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Genes de Plantas , Genes Reporter , Proteínas Luminescentes/química , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Medicago truncatula/genética , Complexos Multiproteicos/química , Complexos Multiproteicos/genética , Complexos Multiproteicos/metabolismo , Floema/química , Floema/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Multimerização Proteica , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Proteína Vermelha Fluorescente
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