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1.
Cell ; 171(2): 287-304.e15, 2017 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-28985561

RESUMO

The evolution of land flora transformed the terrestrial environment. Land plants evolved from an ancestral charophycean alga from which they inherited developmental, biochemical, and cell biological attributes. Additional biochemical and physiological adaptations to land, and a life cycle with an alternation between multicellular haploid and diploid generations that facilitated efficient dispersal of desiccation tolerant spores, evolved in the ancestral land plant. We analyzed the genome of the liverwort Marchantia polymorpha, a member of a basal land plant lineage. Relative to charophycean algae, land plant genomes are characterized by genes encoding novel biochemical pathways, new phytohormone signaling pathways (notably auxin), expanded repertoires of signaling pathways, and increased diversity in some transcription factor families. Compared with other sequenced land plants, M. polymorpha exhibits low genetic redundancy in most regulatory pathways, with this portion of its genome resembling that predicted for the ancestral land plant. PAPERCLIP.


Assuntos
Evolução Biológica , Embriófitas/genética , Genoma de Planta , Marchantia/genética , Adaptação Biológica , Embriófitas/fisiologia , Regulação da Expressão Gênica de Plantas , Marchantia/fisiologia , Anotação de Sequência Molecular , Transdução de Sinais , Transcrição Gênica
2.
Plant Cell ; 34(10): 3512-3542, 2022 09 27.
Artigo em Inglês | MEDLINE | ID: mdl-35976122

RESUMO

The liverwort Marchantia polymorpha has been utilized as a model for biological studies since the 18th century. In the past few decades, there has been a Renaissance in its utilization in genomic and genetic approaches to investigating physiological, developmental, and evolutionary aspects of land plant biology. The reasons for its adoption are similar to those of other genetic models, e.g. simple cultivation, ready access via its worldwide distribution, ease of crossing, facile genetics, and more recently, efficient transformation, genome editing, and genomic resources. The haploid gametophyte dominant life cycle of M. polymorpha is conducive to forward genetic approaches. The lack of ancient whole-genome duplications within liverworts facilitates reverse genetic approaches, and possibly related to this genomic stability, liverworts possess sex chromosomes that evolved in the ancestral liverwort. As a representative of one of the three bryophyte lineages, its phylogenetic position allows comparative approaches to provide insights into ancestral land plants. Given the karyotype and genome stability within liverworts, the resources developed for M. polymorpha have facilitated the development of related species as models for biological processes lacking in M. polymorpha.


Assuntos
Embriófitas , Marchantia , Evolução Biológica , Células Germinativas Vegetais , Marchantia/genética , Filogenia
3.
New Phytol ; 243(4): 1406-1423, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38922903

RESUMO

The GOLDEN2-LIKE (GLK) transcription factors act as a central regulatory node involved in both developmental processes and environmental responses. Marchantia polymorpha, a basal terrestrial plant with strategic evolutionary position, contains a single GLK representative that possesses an additional domain compared to spermatophytes. We analyzed the role of MpGLK in chloroplast biogenesis and development by altering its levels, preforming transcriptomic profiling and conducting chromatin immunoprecipitation. Decreased MpGLK levels impair chloroplast differentiation and disrupt the expression of photosynthesis-associated nuclear genes, while overexpressing MpGLK leads to ectopic chloroplast biogenesis. This demonstrates the MpGLK functions as a bona fide GLK protein, likely representing an ancestral GLK architecture. Altering MpGLK levels directly regulates the expression of genes involved in Chl synthesis and degradation, similar to processes observed in eudicots, and causes various developmental defects in Marchantia, including the formation of dorsal structures such as air pores and gemma cups. MpGLK, also directly activates MpMAX2 gene expression, regulating the timing of gemma cup development. Our study shows that MpGLK functions as a master regulator, potentially coupling chloroplast development with vegetative reproduction. This illustrates the complex regulatory networks governing chloroplast function and plant development communication and highlight the evolutionary conservation of GLK-mediated regulatory processes across plant species.


Assuntos
Cloroplastos , Regulação da Expressão Gênica de Plantas , Marchantia , Proteínas de Plantas , Fatores de Transcrição , Marchantia/genética , Marchantia/crescimento & desenvolvimento , Marchantia/metabolismo , Cloroplastos/metabolismo , Cloroplastos/genética , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Desenvolvimento Vegetal/genética , Fotossíntese/genética
4.
Bull Entomol Res ; 114(4): 563-570, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39295441

RESUMO

Confronting environments with low relative humidity is one of the main challenges faced by insects with expanding distribution ranges. Anastrepha ludens (the Mexican fruit fly) has evolved to cope with the variable conditions encountered during its lifetime, which allows it to colonise a wide range of environments. However, our understanding of the mechanisms underpinning the ability of this species to confront environments with low relative humidity is incomplete. In this sense, omic approaches such as transcriptomics can be helpful for advancing our knowledge on how this species copes with desiccation stress. Considering this, in this study, we performed transcriptomic analyses to compare the molecular responses of laboratory-reared A. ludens exposed and unexposed to desiccation. Data from the transcriptome analyses indicated that the responses to desiccation are shared by both sexes. We identified the up-regulation of transcripts encoding proteins involved in lipid metabolism and membrane remodelling, as well as proteases and cuticular proteins. Our results provide a framework for understanding the response to desiccation stress in one of the most invasive fruit fly species in the world.


Assuntos
Dessecação , Tephritidae , Animais , Tephritidae/genética , Tephritidae/fisiologia , Feminino , Masculino , Transcriptoma , Perfilação da Expressão Gênica , Estresse Fisiológico
5.
Plant J ; 109(4): 873-890, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34807478

RESUMO

Trichoderma atroviride is a root-colonizing fungus that confers multiple benefits to plants. In plants, small RNA (sRNA)-mediated gene silencing (sRNA-MGS) plays pivotal roles in growth, development, and pathogen attack. Here, we explored the role of core components of Arabidopsis thaliana sRNA-MGS pathways during its interaction with Trichoderma. Upon interaction with Trichoderma, sRNA-MGS-related genes paralleled the expression of Arabidopsis defense-related genes, linked to salicylic acid (SA) and jasmonic acid (JA) pathways. SA- and JA-related genes were primed by Trichoderma in leaves after the application of the well-known pathogen-associated molecular patterns flg22 and chitin, respectively. Defense-related genes were primed in roots as well, but to different extents and behaviors. Phenotypical characterization of mutants in AGO genes and components of the RNA-dependent DNA methylation (RdDM) pathway revealed that different sets of sRNA-MGS-related genes are essential for (i) the induction of systemic acquired resistance against Botrytis cinerea, (ii) the activation of the expression of plant defense-related genes, and (iii) root colonization by Trichoderma. Additionally, plant growth induced by Trichoderma depends on functional RdDM. Profiling of DNA methylation and histone N-tail modification patterns at the Arabidopsis Nitrile-Specifier Protein-4 (NSP4) locus, which is responsive to Trichoderma, showed altered epigenetic modifications in RdDM mutants. Furthermore, NSP4 is required for the induction of systemic acquired resistance against Botrytis and avoidance of enhanced root colonization by Trichoderma. Together, our results indicate that RdDM is essential in Arabidopsis to establish a beneficial relationship with Trichoderma. We propose that DNA methylation and histone modifications are required for plant priming by the beneficial fungus against B. cinerea.


Assuntos
Arabidopsis/genética , Arabidopsis/metabolismo , Resistência à Doença/genética , Inativação Gênica , Hypocreales/genética , Nitrilas/metabolismo , RNA/metabolismo , Proteínas de Arabidopsis/metabolismo , Botrytis , Ciclopentanos , Regulação da Expressão Gênica de Plantas , Hypocreales/metabolismo , Oxilipinas , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Raízes de Plantas/metabolismo , Ácido Salicílico/metabolismo , Trichoderma/genética , Trichoderma/metabolismo
6.
Planta ; 257(6): 105, 2023 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-37120771

RESUMO

MAIN CONCLUSION: Our study presents evidence for a novel mechanism for RBR function in transcriptional gene silencing by interacting with key players of the RdDM pathway in Arabidopsis and several plant clades. Transposable elements and other repetitive elements are silenced by the RNA-directed DNA methylation pathway (RdDM). In RdDM, POLIV-derived transcripts are converted into double-stranded RNA (dsRNA) by the activity of RDR2 and subsequently processed into 24 nucleotide short interfering RNAs (24-nt siRNAs) by DCL3. 24-nt siRNAs serve as guides to direct AGO4-siRNA complexes to chromatin-bound POLV-derived transcripts generated from the template/target DNA. The interaction between POLV, AGO4, DMS3, DRD1, RDM1 and DRM2 promotes DRM2-mediated de novo DNA methylation. The Arabidopsis Retinoblastoma protein homolog (RBR) is a master regulator of the cell cycle, stem cell maintenance, and development. We in silico predicted and explored experimentally the protein-protein interactions (PPIs) between RBR and members of the RdDM pathway. We found that the largest subunits of POLIV and POLV (NRPD1 and NRPE1), the shared second largest subunit of POLIV and POLV (NRPD/E2), RDR1, RDR2, DCL3, DRM2, and SUVR2 contain canonical and non-canonical RBR binding motifs and several of them are conserved since algae and bryophytes. We validated experimentally PPIs between Arabidopsis RBR and several of the RdDM pathway proteins. Moreover, seedlings from loss-of-function mutants in RdDM and RBR show similar phenotypes in the root apical meristem. We show that RdDM and SUVR2 targets are up-regulated in the 35S:AmiGO-RBR background.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Neoplasias da Retina , Retinoblastoma , Arabidopsis/genética , Arabidopsis/metabolismo , Metilação de DNA/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , RNA Polimerases Dirigidas por DNA/genética , Retinoblastoma/genética , RNA Interferente Pequeno/genética , RNA de Cadeia Dupla/metabolismo , RNA de Plantas/genética , RNA de Plantas/metabolismo , Neoplasias da Retina/genética , Regulação da Expressão Gênica de Plantas , Ribonuclease III/genética
7.
Int J Mol Sci ; 21(21)2020 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-33171770

RESUMO

Phosphate (Pi) is a pivotal nutrient that constraints plant development and productivity in natural ecosystems. Land colonization by plants, more than 470 million years ago, evolved adaptive mechanisms to conquer Pi-scarce environments. However, little is known about the molecular basis underlying such adaptations at early branches of plant phylogeny. To shed light on how early divergent plants respond to Pi limitation, we analyzed the morpho-physiological and transcriptional dynamics of Marchantia polymorpha upon Pi starvation. Our phylogenomic analysis highlights some gene networks present since the Chlorophytes and others established in the Streptophytes (e.g., PHR1-SPX1 and STOP1-ALMT1, respectively). At the morpho-physiological level, the response is characterized by the induction of phosphatase activity, media acidification, accumulation of auronidins, reduction of internal Pi concentration, and developmental modifications of rhizoids. The transcriptional response involves the induction of MpPHR1, Pi transporters, lipid turnover enzymes, and MpMYB14, which is an essential transcription factor for auronidins biosynthesis. MpSTOP2 up-regulation correlates with expression changes in genes related to organic acid biosynthesis and transport, suggesting a preference for citrate exudation. An analysis of MpPHR1 binding sequences (P1BS) shows an enrichment of this cis regulatory element in differentially expressed genes. Our study unravels the strategies, at diverse levels of organization, exerted by M. polymorpha to cope with low Pi availability.


Assuntos
Marchantia/genética , Marchantia/metabolismo , Fosfatos/metabolismo , Ecossistema , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/genética , Redes Reguladoras de Genes/efeitos dos fármacos , Redes Reguladoras de Genes/genética , Hepatófitas/metabolismo , Filogenia , Fatores de Transcrição/metabolismo
8.
Dev Biol ; 433(2): 227-239, 2018 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-29291975

RESUMO

The axolotl (Ambystoma mexicanum) is the vertebrate model system with the highest regeneration capacity. Experimental tools established over the past 100 years have been fundamental to start unraveling the cellular and molecular basis of tissue and limb regeneration. In the absence of a reference genome for the Axolotl, transcriptomic analysis become fundamental to understand the genetic basis of regeneration. Here we present one of the most diverse transcriptomic data sets for Axolotl by profiling coding and non-coding RNAs from diverse tissues. We reconstructed a population of 115,906 putative protein coding mRNAs as full ORFs (including isoforms). We also identified 352 conserved miRNAs and 297 novel putative mature miRNAs. Systematic enrichment analysis of gene expression allowed us to identify tissue-specific protein-coding transcripts. We also found putative novel and conserved microRNAs which potentially target mRNAs which are reported as important disease candidates in heart and liver.


Assuntos
Ambystoma mexicanum/genética , Regulação da Expressão Gênica , RNA Mensageiro/genética , Regeneração/genética , Transcrição Gênica , Transcriptoma , Ambystoma mexicanum/fisiologia , Animais , Feminino , Biblioteca Gênica , Ontologia Genética , Humanos , MicroRNAs/biossíntese , MicroRNAs/genética , Especificidade de Órgãos , Análise de Componente Principal , RNA Mensageiro/biossíntese , RNA Interferente Pequeno/genética , Análise de Sequência de RNA , Especificidade da Espécie
9.
New Phytol ; 223(2): 575-581, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30920664

RESUMO

Methylation of DNA is an epigenetic mechanism for the control of gene expression. Alterations in the regulatory pathways involved in the establishment, perpetuation and removal of DNA methylation can lead to severe developmental alterations. Our understanding of the mechanistic aspects and relevance of DNA methylation comes from remarkable studies in well-established angiosperm plant models including maize and Arabidopsis. The study of plant models positioned at basal lineages opens exciting opportunities to expand our knowledge on the function and evolution of the components of DNA methylation. In this Tansley Insight, we summarize current progress in our understanding of the molecular basis and relevance of DNA methylation in the liverwort Marchantia polymorpha.


Assuntos
Metilação de DNA/genética , Marchantia/genética , RNA Polimerases Dirigidas por DNA/metabolismo , Marchantia/crescimento & desenvolvimento , Modelos Biológicos , RNA de Plantas/metabolismo
10.
Nature ; 498(7452): 94-8, 2013 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-23665961

RESUMO

It has been argued that the evolution of plant genome size is principally unidirectional and increasing owing to the varied action of whole-genome duplications (WGDs) and mobile element proliferation. However, extreme genome size reductions have been reported in the angiosperm family tree. Here we report the sequence of the 82-megabase genome of the carnivorous bladderwort plant Utricularia gibba. Despite its tiny size, the U. gibba genome accommodates a typical number of genes for a plant, with the main difference from other plant genomes arising from a drastic reduction in non-genic DNA. Unexpectedly, we identified at least three rounds of WGD in U. gibba since common ancestry with tomato (Solanum) and grape (Vitis). The compressed architecture of the U. gibba genome indicates that a small fraction of intergenic DNA, with few or no active retrotransposons, is sufficient to regulate and integrate all the processes required for the development and reproduction of a complex organism.


Assuntos
Evolução Molecular , Genoma de Planta/genética , Magnoliopsida/genética , DNA Intergênico/genética , Duplicação Gênica/genética , Genes de Plantas/genética , Modelos Genéticos , Solanum/genética , Sintenia/genética , Vitis/genética
11.
Plant Cell Physiol ; 59(12): 2421-2431, 2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30102384

RESUMO

DNA methylation is an epigenetic mark that ensures silencing of transposable elements (TEs) and affects gene expression in many organisms. The function of different DNA methylation regulatory pathways has been largely characterized in the model plant Arabidopsis thaliana. However, far less is known about DNA methylation regulation and functions in basal land plants. Here we focus on the liverwort Marchantia polymorpha, an emerging model species that represents a basal lineage of land plants. We identified MpMET, the M. polymorpha ortholog of the METHYLTRANSFERASE 1 (MET1) gene required for maintenance of methylation at CG sites in angiosperms. We generated Mpmet mutants using the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein9) system, which showed a significant loss of CG methylation and severe morphological changes and developmental defects. The mutants developed many adventitious shoot-like structures, suggesting that MpMET is required for maintaining differentiated cellular identities in the gametophyte. Even though numerous TEs were up-regulated, non-CG methylation was generally highly increased at TEs in the Mpmet mutants. Closer inspection of CHG methylation revealed features unique to M. polymorpha. Methylation of CCG sites in M. polymorpha does not depend on MET1, unlike in A. thaliana and Physcomitrella patens. Our results highlight the diversity of non-CG methylation regulatory mechanisms in plants.


Assuntos
Divisão Celular/genética , Ilhas de CpG/genética , Metilação de DNA/genética , Marchantia/citologia , Marchantia/genética , Elementos de DNA Transponíveis/genética , Genoma de Planta , Mutação/genética
12.
Mol Biol Evol ; 32(7): 1788-99, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25767205

RESUMO

Small RNA-mediated chromatin modification is a conserved feature of eukaryotes. In flowering plants, the short interfering (si)RNAs that direct transcriptional silencing are abundant and subfunctionalization has led to specialized machinery responsible for synthesis and action of these small RNAs. In particular, plants possess polymerase (Pol) IV and Pol V, multi-subunit homologs of the canonical DNA-dependent RNA Pol II, as well as specialized members of the RNA-dependent RNA Polymerase (RDR), Dicer-like (DCL), and Argonaute (AGO) families. Together these enzymes are required for production and activity of Pol IV-dependent (p4-)siRNAs, which trigger RNA-directed DNA methylation (RdDM) at homologous sequences. p4-siRNAs accumulate highly in developing endosperm, a specialized tissue found only in flowering plants, and are rare in nonflowering plants, suggesting that the evolution of flowers might coincide with the emergence of specialized RdDM machinery. Through comprehensive identification of RdDM genes from species representing the breadth of the land plant phylogeny, we describe the ancient origin of Pol IV and Pol V, suggesting that a nearly complete and functional RdDM pathway could have existed in the earliest land plants. We also uncover innovations in these enzymes that are coincident with the emergence of seed plants and flowering plants, and recent duplications that might indicate additional subfunctionalization. Phylogenetic analysis reveals rapid evolution of Pol IV and Pol V subunits relative to their Pol II counterparts and suggests that duplicates were retained and subfunctionalized through Escape from Adaptive Conflict. Evolution within the carboxy-terminal domain of the Pol V largest subunit is particularly striking, where illegitimate recombination facilitated extreme sequence divergence.


Assuntos
RNA Polimerases Dirigidas por DNA/genética , Filogenia , Proteínas de Plantas/genética , Plantas/enzimologia , Plantas/genética , Sequência de Aminoácidos , RNA Polimerases Dirigidas por DNA/química , Evolução Molecular , Flores/genética , Duplicação Gênica , Inativação Gênica , Genes de Plantas , Magnoliopsida/enzimologia , Dados de Sequência Molecular , Proteínas de Plantas/química , Estrutura Terciária de Proteína , Subunidades Proteicas/genética , Especificidade da Espécie
13.
Plant Cell Physiol ; 57(2): 339-58, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26861787

RESUMO

Bryophytes (liverworts, hornworts and mosses) comprise the three earliest diverging lineages of land plants (embryophytes). Marchantia polymorpha, a complex thalloid Marchantiopsida liverwort that has been developed into a model genetic system, occupies a key phylogenetic position. Therefore, M. polymorpha is useful in studies aiming to elucidate the evolution of gene regulation mechanisms in plants. In this study, we used computational, transcriptomic, small RNA and degradome analyses to characterize microRNA (miRNA)-mediated pathways of gene regulation in M. polymorpha. The data have been integrated into the open access ContigViews-miRNA platform for further reference. In addition to core components of the miRNA pathway, 129 unique miRNA sequences, 11 of which could be classified into seven miRNA families that are conserved in embryophytes (miR166a, miR390, miR529c, miR171-3p, miR408a, miR160 and miR319a), were identified. A combination of computational and degradome analyses allowed us to identify and experimentally validate 249 targets. In some cases, the target genes are orthologous to those of other embryophytes, but in other cases, the conserved miRNAs target either paralogs or members of different gene families. In addition, the newly discovered Mpo-miR11707.1 and Mpo-miR11707.2 are generated from a common precursor and target MpARGONAUTE1 (LW1759). Two other newly discovered miRNAs, Mpo-miR11687.1 and Mpo-miR11681.1, target the MADS-box transcription factors MpMADS1 and MpMADS2, respectively. Interestingly, one of the pentatricopeptide repeat (PPR) gene family members, MpPPR_66 (LW9825), the protein products of which are generally involved in various steps of RNA metabolism, has a long stem-loop transcript that can generate Mpo-miR11692.1 to autoregulate MpPPR_66 (LW9825) mRNA. This study provides a foundation for further investigations of the RNA-mediated silencing mechanism in M. polymorpha as well as of the evolution of this gene silencing pathway in embryophytes.


Assuntos
Marchantia/genética , MicroRNAs/genética , Estabilidade de RNA/genética , Análise de Sequência de RNA/métodos , Sequência de Bases , Sequência Conservada/genética , Regulação para Baixo/genética , Perfilação da Expressão Gênica , Inativação Gênica , Genes de Plantas , Genes Reporter , MicroRNAs/metabolismo , Anotação de Sequência Molecular , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Filogenia , Transcriptoma/genética
14.
Plant Cell Physiol ; 57(2): 257-61, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26644462

RESUMO

While Marchantia polymorpha has been utilized as a model system to investigate fundamental biological questions for over almost two centuries, there is renewed interest in M. polymorpha as a model genetic organism in the genomics era. Here we outline community guidelines for M. polymorpha gene and transgene nomenclature, and we anticipate that these guidelines will promote consistency and reduce both redundancy and confusion in the scientific literature.


Assuntos
Genes de Plantas , Guias como Assunto , Marchantia/classificação , Marchantia/genética , Terminologia como Assunto , Transgenes
15.
PLoS Biol ; 11(11): e1001724, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24302889

RESUMO

Quiescent long-term somatic stem cells reside in plant and animal stem cell niches. Within the Arabidopsis root stem cell population, the Quiescent Centre (QC), which contains slowly dividing cells, maintains surrounding short-term stem cells and may act as a long-term reservoir for stem cells. The RETINOBLASTOMA-RELATED (RBR) protein cell-autonomously reinforces mitotic quiescence in the QC. RBR interacts with the stem cell transcription factor SCARECROW (SCR) through an LxCxE motif. Disruption of this interaction by point mutation in SCR or RBR promotes asymmetric divisions in the QC that renew short-term stem cells. Analysis of the in vivo role of quiescence in the root stem cell niche reveals that slow cycling within the QC is not needed for structural integrity of the niche but allows the growing root to cope with DNA damage.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/citologia , Meristema/citologia , Sequência de Aminoácidos , Arabidopsis/metabolismo , Proteínas de Arabidopsis/química , Proliferação de Células , Técnicas de Silenciamento de Genes , Meristema/metabolismo , Ligação Proteica , Domínios e Motivos de Interação entre Proteínas , Mapas de Interação de Proteínas , Nicho de Células-Tronco , Células-Tronco/fisiologia
16.
Nature ; 464(7288): 628-32, 2010 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-20208518

RESUMO

In the ovules of most sexual flowering plants female gametogenesis is initiated from a single surviving gametic cell, the functional megaspore, formed after meiosis of the somatically derived megaspore mother cell (MMC). Because some mutants and certain sexual species exhibit more than one MMC, and many others are able to form gametes without meiosis (by apomixis), it has been suggested that somatic cells in the ovule are competent to respond to a local signal likely to have an important function in determination. Here we show that the Arabidopsis protein ARGONAUTE 9 (AGO9) controls female gamete formation by restricting the specification of gametophyte precursors in a dosage-dependent, non-cell-autonomous manner. Mutations in AGO9 lead to the differentiation of multiple gametic cells that are able to initiate gametogenesis. The AGO9 protein is not expressed in the gamete lineage; instead, it is expressed in cytoplasmic foci of somatic companion cells. Mutations in SUPPRESSOR OF GENE SILENCING 3 and RNA-DEPENDENT RNA POLYMERASE 6 exhibit an identical defect to ago9 mutants, indicating that the movement of small RNA (sRNAs) silencing out of somatic companion cells is necessary for controlling the specification of gametic cells. AGO9 preferentially interacts with 24-nucleotide sRNAs derived from transposable elements (TEs), and its activity is necessary to silence TEs in female gametes and their accessory cells. Our results show that AGO9-dependent sRNA silencing is crucial to specify cell fate in the Arabidopsis ovule, and that epigenetic reprogramming in companion cells is necessary for sRNA-dependent silencing in plant gametes.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Gametogênese Vegetal/fisiologia , Óvulo Vegetal/metabolismo , RNA de Plantas/metabolismo , Proteínas de Ligação a RNA/metabolismo , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Proteínas Argonautas , Elementos de DNA Transponíveis/genética , Regulação da Expressão Gênica de Plantas , Inativação Gênica , Meiose , Dados de Sequência Molecular , Mutagênese Insercional/genética , Óvulo Vegetal/crescimento & desenvolvimento , Fenótipo , Proteínas de Ligação a RNA/genética
17.
PLoS Genet ; 9(10): e1003773, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24146624

RESUMO

Paramutation is a well-studied epigenetic phenomenon in which trans communication between two different alleles leads to meiotically heritable transcriptional silencing of one of the alleles. Paramutation at the b1 locus involves RNA-mediated transcriptional silencing and requires specific tandem repeats that generate siRNAs. This study addressed three important questions: 1) are the tandem repeats sufficient for paramutation, 2) do they need to be in an allelic position to mediate paramutation, and 3) is there an association between the ability to mediate paramutation and repeat DNA methylation levels? Paramutation was achieved using multiple transgenes containing the b1 tandem repeats, including events with tandem repeats of only one half of the repeat unit (413 bp), demonstrating that these sequences are sufficient for paramutation and an allelic position is not required for the repeats to communicate. Furthermore, the transgenic tandem repeats increased the expression of a reporter gene in maize, demonstrating the repeats contain transcriptional regulatory sequences. Transgene-mediated paramutation required the mediator of paramutation1 gene, which is necessary for endogenous paramutation, suggesting endogenous and transgene-mediated paramutation both require an RNA-mediated transcriptional silencing pathway. While all tested repeat transgenes produced small interfering RNAs (siRNAs), not all transgenes induced paramutation suggesting that, as with endogenous alleles, siRNA production is not sufficient for paramutation. The repeat transgene-induced silencing was less efficiently transmitted than silencing induced by the repeats of endogenous b1 alleles, which is always 100% efficient. The variability in the strength of the repeat transgene-induced silencing enabled testing whether the extent of DNA methylation within the repeats correlated with differences in efficiency of paramutation. Transgene-induced paramutation does not require extensive DNA methylation within the transgene. However, increased DNA methylation within the endogenous b1 repeats after transgene-induced paramutation was associated with stronger silencing of the endogenous allele.


Assuntos
Metilação de DNA/genética , Epigênese Genética , Sequências de Repetição em Tandem/genética , Transcrição Gênica , Alelos , Sequência de Bases , Regulação da Expressão Gênica de Plantas , Inativação Gênica , Mutação , Plantas Geneticamente Modificadas/genética , RNA/genética , RNA Interferente Pequeno/genética , Zea mays/genética , Zea mays/metabolismo
18.
PLoS One ; 19(6): e0304790, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38875250

RESUMO

In plants, small RNAs (sRNAs), mainly microRNAs (miRNAs) and small interfering RNAs (siRNAs), have been described as key regulators of plant development, growth, and abiotic and biotic responses. Despite reports indicating the involvement of certain sRNAs in regulating the interaction between Botrytis cinerea (a major necrotrophic fungal phytopathogen) and host plants, there remains a lack of analysis regarding the potential regulatory roles of plant sRNAs during early stages of the interaction despite early immune responses observed then during infection. We present the first transcriptome-wide analysis of small RNA expression on the early interaction between the necrotrophic fungus Botrytis cinerea and the model plant Arabidopsis thaliana. We found that evolutionary conserved A. thaliana miRNAs were the sRNAs that accumulated the most in the presence of B. cinerea. The upregulation of miR167, miR159 and miR319 was of particular interest because these, together with their target transcripts, are involved in the fine regulation of the plant hormone signaling pathways. We also describe that miR173, which triggers the production of secondary siRNAs from TAS1 and TAS2 loci, as well as secondary siRNAs derived from these loci, is upregulated in response to B. cinerea. Thus, at an early stage of the interaction there are transcriptional changes of sRNA-guided silencing pathway genes and of a subset of sRNAs that targeted genes from the PPR gene superfamily, and these may be important mechanisms regulating the interaction between A. thaliana and B. cinerea. This work provides the basis for a better understanding of the regulation mediated by sRNAs during early B. cinerea-plant interaction and may help in the development of more effective strategies for its control.


Assuntos
Arabidopsis , Botrytis , Regulação da Expressão Gênica de Plantas , Interações Hospedeiro-Patógeno , MicroRNAs , RNA de Plantas , Botrytis/genética , Botrytis/patogenicidade , Arabidopsis/genética , Arabidopsis/microbiologia , MicroRNAs/genética , MicroRNAs/metabolismo , Interações Hospedeiro-Patógeno/genética , RNA de Plantas/genética , Doenças das Plantas/microbiologia , Doenças das Plantas/genética , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Perfilação da Expressão Gênica
19.
Proc Natl Acad Sci U S A ; 107(29): 12986-91, 2010 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-20616013

RESUMO

Paramutation is the epigenetic transfer of information between alleles that leads to the heritable change of expression of one allele. Paramutation at the b1 locus in maize requires seven noncoding tandem repeat (b1TR) sequences located approximately 100 kb upstream of the transcription start site of b1, and mutations in several genes required for paramutation implicate an RNA-mediated mechanism. The mediator of paramutation (mop1) gene, which encodes a protein closely related to RNA-dependent RNA polymerases, is absolutely required for paramutation. Herein, we investigate the potential function of mop1 and the siRNAs that are produced from the b1TR sequences. Production of siRNAs from the b1TR sequences depends on a functional mop1 gene, but transcription of the repeats is not dependent on mop1. Further nuclear transcription assays suggest that the b1TR sequences are likely transcribed predominantly by RNA polymerase II. To address whether production of b1TR-siRNAs correlated with paramutation, we examined siRNA production in alleles that cannot undergo paramutation. Alleles that cannot participate in paramutation also produce b1TR-siRNAs, suggesting that b1TR-siRNAs are not sufficient for paramutation in the tissues analyzed. However, when b1TR-siRNAs are produced from a transgene expressing a hairpin RNA, b1 paramutation can be recapitulated. We hypothesize that either the b1TR-siRNAs or the dsRNA template mediates the trans-communication between the alleles that establishes paramutation. In addition, we uncovered a role for mop1 in the biogenesis of a subset of microRNAs (miRNAs) and show that it functions at the level of production of the primary miRNA transcripts.


Assuntos
Loci Gênicos/genética , Mutação/genética , RNA de Plantas/metabolismo , Zea mays/genética , Alelos , Sequência de Bases , RNA Polimerases Dirigidas por DNA/metabolismo , MicroRNAs/biossíntese , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , RNA de Cadeia Dupla/biossíntese , RNA Interferente Pequeno/biossíntese , Sequências de Repetição em Tandem/genética , Transcrição Gênica , Transgenes/genética
20.
PeerJ ; 11: e14581, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36632141

RESUMO

Dragon fruit, pitahaya or pitaya are common names for the species in the Hylocereus group of Selenicereus that produce edible fruit. These Neotropical epiphytic cacti are considered promising underutilized crops and are currently cultivated around the world. The most important species, S. undatus, has been managed in the Maya domain for centuries and is the focus of this article. Transcriptome profiles from stems of wild and cultivated plants of this species were compared. We hypothesized that differences in transcriptomic signatures could be associated with genes related to drought stress. De novo transcriptome assembly and the analysis of differentially expressed genes (DEGs) allowed us to identify a total of 9,203 DEGs in the Hunucmá cultivar relative of wild Mozomboa plants. Of these, 4,883 represent up-regulated genes and 4,320, down-regulated genes. Additionally, 6,568 DEGs were identified from a comparison between the Umán cultivar and wild plants, revealing 3,286 up-regulated and 3,282 down-regulated genes. Approximately half of the DEGs are shared by the two cultivated plants. Differences between the two cultivars that were collected in the same region could be the result of differences in management. Metabolism was the most representative functional category in both cultivars. The up-regulated genes of both cultivars formed a network related to the hormone-mediated signaling pathway that includes cellular responses to auxin stimulus and to hormone stimulus. These cellular reactions have been documented in several cultivated plants in which drought-tolerant cultivars modify auxin transport and ethylene signaling, resulting in a better redistribution of assimilates.


Assuntos
Cactaceae , Transcriptoma , Transcriptoma/genética , Frutas/genética , Cactaceae/genética , Ácidos Indolacéticos , Hormônios
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