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1.
PLoS Genet ; 19(7): e1010344, 2023 07.
Article in English | MEDLINE | ID: mdl-37418499

ABSTRACT

The chloroplast proteome is a dynamic mosaic of plastid- and nuclear-encoded proteins. Plastid protein homeostasis is maintained through the balance between de novo synthesis and proteolysis. Intracellular communication pathways, including the plastid-to-nucleus signalling and the protein homeostasis machinery, made of stromal chaperones and proteases, shape chloroplast proteome based on developmental and physiological needs. However, the maintenance of fully functional chloroplasts is costly and under specific stress conditions the degradation of damaged chloroplasts is essential to the maintenance of a healthy population of photosynthesising organelles while promoting nutrient redistribution to sink tissues. In this work, we have addressed this complex regulatory chloroplast-quality-control pathway by modulating the expression of two nuclear genes encoding plastid ribosomal proteins PRPS1 and PRPL4. By transcriptomics, proteomics and transmission electron microscopy analyses, we show that the increased expression of PRPS1 gene leads to chloroplast degradation and early flowering, as an escape strategy from stress. On the contrary, the overaccumulation of PRPL4 protein is kept under control by increasing the amount of plastid chaperones and components of the unfolded protein response (cpUPR) regulatory mechanism. This study advances our understanding of molecular mechanisms underlying chloroplast retrograde communication and provides new insights into cellular responses to impaired plastid protein homeostasis.


Subject(s)
Proteome , Proteostasis , Proteostasis/genetics , Proteome/genetics , Proteome/metabolism , Plastids/genetics , Plastids/metabolism , Chloroplasts/genetics , Chloroplasts/metabolism , Signal Transduction/physiology , Chloroplast Proteins/metabolism , Gene Expression Regulation, Plant
2.
Plant Physiol ; 194(4): 2117-2135, 2024 Mar 29.
Article in English | MEDLINE | ID: mdl-38060625

ABSTRACT

The gynoecium is critical for the reproduction of flowering plants as it contains the ovules and the tissues that foster pollen germination, growth, and guidance. These tissues, known as the reproductive tract (ReT), comprise the stigma, style, and transmitting tract (TT). The ReT and ovules originate from the carpel margin meristem (CMM) within the pistil. SHOOT MERISTEMLESS (STM) is a key transcription factor for meristem formation and maintenance. In all above-ground meristems, including the CMM, local STM downregulation is required for organ formation. However, how this downregulation is achieved in the CMM is unknown. Here, we have studied the role of HISTONE DEACETYLASE 19 (HDA19) in Arabidopsis (Arabidopsis thaliana) during ovule and ReT differentiation based on the observation that the hda19-3 mutant displays a reduced ovule number and fails to differentiate the TT properly. Fluorescence-activated cell sorting coupled with RNA-sequencing revealed that in the CMM of hda19-3 mutants, genes promoting organ development are downregulated while meristematic markers, including STM, are upregulated. HDA19 was essential to downregulate STM in the CMM, thereby allowing ovule formation and TT differentiation. STM is ectopically expressed in hda19-3 at intermediate stages of pistil development, and its downregulation by RNA interference alleviated the hda19-3 phenotype. Chromatin immunoprecipitation assays indicated that STM is a direct target of HDA19 during pistil development and that the transcription factor SEEDSTICK is also required to regulate STM via histone acetylation. Thus, we identified factors required for the downregulation of STM in the CMM, which is necessary for organogenesis and tissue differentiation.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Histones/genetics , Ovule/genetics , Ovule/metabolism , Arabidopsis/physiology , Transcription Factors/metabolism , Meristem , Gene Expression Regulation, Plant , MADS Domain Proteins/genetics , Histone Deacetylases/metabolism
3.
Cell ; 132(2): 273-85, 2008 Jan 25.
Article in English | MEDLINE | ID: mdl-18243102

ABSTRACT

During photosynthesis, two photoreaction centers located in the thylakoid membranes of the chloroplast, photosystems I and II (PSI and PSII), use light energy to mobilize electrons to generate ATP and NADPH. Different modes of electron flow exist, of which the linear electron flow is driven by PSI and PSII, generating ATP and NADPH, whereas the cyclic electron flow (CEF) only generates ATP and is driven by the PSI alone. Different environmental and metabolic conditions require the adjustment of ATP/NADPH ratios and a switch of electron distribution between the two photosystems. With the exception of PGR5, other components facilitating CEF are unknown. Here, we report the identification of PGRL1, a transmembrane protein present in thylakoids of Arabidopsis thaliana. Plants lacking PGRL1 show perturbation of CEF, similar to PGR5-deficient plants. We find that PGRL1 and PGR5 interact physically and associate with PSI. We therefore propose that the PGRL1-PGR5 complex facilitates CEF in eukaryotes.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Membrane Proteins/metabolism , Photosynthetic Reaction Center Complex Proteins/metabolism , Photosystem I Protein Complex/metabolism , Thylakoids/chemistry , Adenosine Triphosphate/biosynthesis , Amino Acid Sequence , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis Proteins/chemistry , Arabidopsis Proteins/genetics , Chloroplasts/metabolism , DNA, Plant/genetics , DNA, Plant/isolation & purification , Electron Transport , Gene Expression Regulation, Plant , Genes, Plant , Kinetics , Membrane Proteins/chemistry , Membrane Proteins/genetics , Models, Biological , Molecular Sequence Data , Mutation , NADP/biosynthesis , Oxidation-Reduction , Photosynthetic Reaction Center Complex Proteins/genetics , Plastoquinone/metabolism , Protein Isoforms/chemistry , Protein Isoforms/genetics , Protein Isoforms/metabolism , Protein Structure, Secondary , Protein Structure, Tertiary , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/metabolism , Sequence Homology, Amino Acid , Subcellular Fractions/metabolism
4.
BMC Plant Biol ; 21(1): 238, 2021 May 27.
Article in English | MEDLINE | ID: mdl-34044765

ABSTRACT

Fruits and seeds resulting from fertilization of flowers, represent an incredible evolutionary advantage in angiosperms and have seen them become a critical element in our food supply.Many studies have been conducted to reveal how fruit matures while protecting growing seeds and ensuring their dispersal. As result, several transcription factors involved in fruit maturation and senescence have been isolated both in model and crop plants. These regulators modulate several cellular processes that occur during fruit ripening such as chlorophyll breakdown, tissue softening, carbohydrates and pigments accumulation.The NAC superfamily of transcription factors is known to be involved in almost all these aspects of fruit development and maturation. In this review, we summarise the current knowledge regarding NACs that modulate fruit ripening in model species (Arabidopsis thaliana and Solanum lycopersicum) and in crops of commercial interest (Oryza sativa, Malus domestica, Fragaria genus, Citrus sinensis and Musa acuminata).


Subject(s)
Arabidopsis/genetics , Fruit/genetics , Solanum lycopersicum/genetics , Transcription Factors/metabolism , Arabidopsis/growth & development , Arabidopsis/physiology , Fruit/enzymology , Fruit/physiology , Solanum lycopersicum/growth & development , Pigmentation , Transcription Factors/genetics
5.
Int J Mol Sci ; 22(5)2021 Mar 05.
Article in English | MEDLINE | ID: mdl-33807566

ABSTRACT

Angiosperm reproduction relies on the precise growth of the pollen tube through different pistil tissues carrying two sperm cells into the ovules' embryo sac, where they fuse with the egg and the central cell to accomplish double fertilization and ultimately initiate seed development. A network of intrinsic and tightly regulated communication and signaling cascades, which mediate continuous interactions between the pollen tube and the sporophytic and gametophytic female tissues, ensures the fast and meticulous growth of pollen tubes along the pistil, until it reaches the ovule embryo sac. Most of the pollen tube growth occurs in a specialized tissue-the transmitting tract-connecting the stigma, the style, and the ovary. This tissue is composed of highly secretory cells responsible for producing an extensive extracellular matrix. This multifaceted matrix is proposed to support and provide nutrition and adhesion for pollen tube growth and guidance. Insights pertaining to the mechanisms that underlie these processes remain sparse due to the difficulty of accessing and manipulating the female sporophytic tissues enclosed in the pistil. Here, we summarize the current knowledge on this key step of reproduction in flowering plants with special emphasis on the female transmitting tract tissue.


Subject(s)
Fertilization/physiology , Flowers/physiology , Ovule/physiology , Extracellular Matrix/physiology , Flowers/metabolism , Magnoliopsida/metabolism , Magnoliopsida/physiology , Ovule/metabolism , Plant Proteins/metabolism , Pollen Tube/metabolism , Pollen Tube/physiology , Seeds/metabolism , Signal Transduction/physiology
6.
Eur Heart J ; 40(36): 3026-3032, 2019 09 21.
Article in English | MEDLINE | ID: mdl-31377776

ABSTRACT

AIMS: It is unknown whether cardioversion of atrial fibrillation causes thromboembolic events or is a risk marker. To assess causality, we examined the temporal pattern of thromboembolism in patients having cardioversion. METHODS AND RESULTS: We studied patients randomized to aspirin or aspirin plus clopidogrel in the ACTIVE trials, comparing the thromboembolic rate in the peri-cardioversion period (30 days before until 30 days after) to the rate during follow-up, remote from cardioversion. Among 962 patients, the 30-day thromboembolic rate remote from cardioversion was 0.16%; while it was 0.73% in the peri-cardioversion period [hazard ratio (HR) 4.1, 95% confidence interval (CI) 2.1-7.9]. The 30-day thromboembolic rates in the periods immediately before and after cardioversion were 0.47% and 0.96%, respectively (HR 2.2, 95% CI 0.7-7.1). Heart failure (HF) hospitalization increased in the peri-cardioversion period (HR 11.5, 95% CI 6.8-19.4). Compared to baseline, the thromboembolic rate in the 30 days following cardioversion was increased both in patients who received oral anticoagulation or a transoesophageal echocardiogram prior to cardioversion (HR 7.9, 95% CI 2.8-22.4) and in those who did not (HR 4.8, 95% CI 1.6-14.9) (interaction P = 0.2); the risk was also increased with successful (HR 4.5; 95% CI 2.0-10.5) and unsuccessful (HR 10.2; 95% CI 2.3-44.9) cardioversion. CONCLUSIONS: Thromboembolic risk increased in the 30 days before cardioversion and persisted until 30 days post-cardioversion, in a pattern similar to HF hospitalization. These data suggest that the increased thromboembolic risk around the time of cardioversion may not be entirely causal, but confounded by the overall clinical deterioration of patients requiring cardioversion.


Subject(s)
Atrial Fibrillation/therapy , Electric Countershock , Platelet Aggregation Inhibitors/therapeutic use , Thromboembolism/epidemiology , Aged , Aspirin/therapeutic use , Clopidogrel/therapeutic use , Female , Follow-Up Studies , Heart Failure/epidemiology , Hospitalization/statistics & numerical data , Humans , Male , Randomized Controlled Trials as Topic , Risk
7.
Development ; 143(18): 3372-81, 2016 09 15.
Article in English | MEDLINE | ID: mdl-27510967

ABSTRACT

Seed dispersal is an essential trait that enables colonization of new favorable habitats, ensuring species survival. In plants with dehiscent fruits, such as Arabidopsis, seed dispersal depends on two processes: the separation of the fruit valves that protect the seeds (fruit dehiscence) and the detachment of the seeds from the funiculus connecting them to the mother plant (seed abscission). The key factors required to establish a proper lignin pattern for fruit dehiscence are SHATTERPROOF 1 and 2 (SHP1 and SHP2). Here, we demonstrate that the SHP-related gene SEEDSTICK (STK) is a key factor required to establish the proper lignin pattern in the seed abscission zone but in an opposite way. We show that STK acts as a repressor of lignin deposition in the seed abscission zone through the direct repression of HECATE3, whereas the SHP proteins promote lignin deposition in the valve margins by activating INDEHISCENT. The interaction of STK with the SEUSS co-repressor determines the difference in the way STK and SHP proteins control the lignification patterns. Despite this difference in the molecular control of lignification during seed abscission and fruit dehiscence, we show that the genetic networks regulating these two developmental pathways are highly conserved.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Fruit/metabolism , Seed Dispersal/physiology , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Basic Helix-Loop-Helix Transcription Factors/genetics , Basic Helix-Loop-Helix Transcription Factors/metabolism , Fruit/physiology , Gene Regulatory Networks/genetics , Gene Regulatory Networks/physiology , MADS Domain Proteins/genetics , MADS Domain Proteins/metabolism , Seed Dispersal/genetics
8.
Plant Physiol ; 178(3): 1249-1268, 2018 11.
Article in English | MEDLINE | ID: mdl-30275057

ABSTRACT

Fruits protect the developing seeds of angiosperms and actively contribute to seed dispersion. Furthermore, fruit and seed development are highly synchronized and require exchange of information between the mother plant and the developing generations. To explore the mechanisms controlling fruit formation and maturation, we performed a transcriptomic analysis on the valve tissue of the Arabidopsis (Arabidopsis thaliana) silique using RNA sequencing. In doing so, we have generated a data set of differentially regulated genes that will help to elucidate the molecular mechanisms that underpin the initial phase of fruit growth and, subsequently, trigger fruit maturation. The robustness of our data set has been tested by functional genomic studies. Using a reverse genetics approach, we selected 10 differentially expressed genes and explored the consequences of their disruption for both silique growth and senescence. We found that genes contained in our data set play essential roles in different stages of silique development and maturation, indicating that our transcriptome-based gene list is a powerful tool for the elucidation of the molecular mechanisms controlling fruit formation in Arabidopsis.


Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Fruit/genetics , Gene Expression Regulation, Plant , Transcriptome , Arabidopsis/growth & development , Arabidopsis Proteins/metabolism , Flowers/genetics , Flowers/growth & development , Fruit/growth & development , Gene Expression Profiling , Gene Expression Regulation, Developmental , Genes, Reporter , Homeostasis , Promoter Regions, Genetic/genetics , Reverse Genetics , Seeds/genetics , Seeds/growth & development , Sequence Analysis, RNA
9.
J Exp Bot ; 70(11): 2993-3006, 2019 06 01.
Article in English | MEDLINE | ID: mdl-30854549

ABSTRACT

Fruits result from complex biological processes that begin soon after fertilization. Among these processes are cell division and expansion, accumulation of secondary metabolites, and an increase in carbohydrate biosynthesis. Later fruit ripening is accomplished by chlorophyll degradation and cell wall lysis. Fruit maturation is an essential step to optimize seed dispersal, and is controlled by a complex network of transcription factors and genetic regulators that are strongly influenced by phytohormones. Abscisic acid (ABA) and ethylene are the major regulators of ripening and senescence in both dry and fleshy fruits, as demonstrated by numerous ripening-defective mutants, effects of exogenous hormone application, and transcriptome analyses. While ethylene is the best characterized player in the final step of a fruit's life, ABA also has a key regulatory role, promoting ethylene production and acting as a stress-related hormone in response to drought and pathogen attack. In this review, we focus on the role of ABA and ethylene in relation to the interconnected biotic and abiotic phenomena that affect ripening and senescence. We integrate and discuss the most recent data available regarding these biological processes, which are crucial for post-harvest fruit conservation and for food safety.


Subject(s)
Cell Wall/metabolism , Fruit/growth & development , Host-Pathogen Interactions/physiology , Plant Growth Regulators/metabolism , Abscisic Acid/metabolism , Bacterial Physiological Phenomena , Ethylenes/metabolism , Fruit/metabolism , Fruit/microbiology , Fungi/physiology
10.
Planta ; 248(1): 257-265, 2018 Jul.
Article in English | MEDLINE | ID: mdl-29687222

ABSTRACT

MAIN CONCLUSION: AtPPR4-mediated trans-splicing of plastid rps12 transcripts is essential for key embryo morphogenetic events such as development of cotyledons, determination of provascular tissue, and organization of the shoot apical meristem (SAM), but not for the formation of the protodermal layer. Members of the pentatricopeptide repeat (PPR) containing protein family have emerged as key regulators of the organelle post-transcriptional processing and to be essential for proper plant embryo development. In this study, we report the functional characterization of the AtPPR4 (At5g04810) gene encoding a plastid nucleoid PPR protein. In-situ hybridization analysis reveals the presence of AtPPR4 transcripts already at the transition stage of embryo development. As a consequence, embryos lacking the AtPPR4 protein arrest their development at the transition/early-heart stages and show defects in the determination of the provascular tissue and organization of SAM. This complex phenotype is due to the specific role of AtPPR4 in the trans-splicing of the plastid rps12 transcripts, as shown by northern and slot-blot hybridizations, and the consequent defect in 70S ribosome accumulation and plastid protein synthesis, in agreement with the role proposed for the maize orthologue, ZmPPR4.


Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/embryology , Plant Proteins/genetics , Plastids/genetics , Seeds/growth & development , Trans-Splicing , Arabidopsis/genetics , Cotyledon/embryology , In Situ Hybridization , Microscopy, Confocal
11.
Plant Physiol ; 173(1): 155-166, 2017 01.
Article in English | MEDLINE | ID: mdl-27920160

ABSTRACT

The EGG CELL1 (EC1) gene family of Arabidopsis (Arabidopsis thaliana) comprises five members that are specifically expressed in the egg cell and redundantly control gamete fusion during double fertilization. We investigated the activity of all five EC1 promoters in promoter-deletion studies and identified SUF4 (SUPPRESSOR OF FRIGIDA4), a C2H2 transcription factor, as a direct regulator of the EC1 gene expression. In particular, we demonstrated that SUF4 binds to all five Arabidopsis EC1 promoters, thus regulating their expression. The down-regulation of SUF4 in homozygous suf4-1 ovules results in reduced EC1 expression and delayed sperm fusion, which can be rescued by expressing SUF4-ß-glucuronidase under the control of the SUF4 promoter. To identify more gene products able to regulate EC1 expression together with SUF4, we performed coexpression studies that led to the identification of MOM1 (MORPHEUS' MOLECULE1), a component of a silencing mechanism that is independent of DNA methylation marks. In mom1-3 ovules, both SUF4 and EC1 genes are down-regulated, and EC1 genes show higher levels of histone 3 lysine-9 acetylation, suggesting that MOM1 contributes to the regulation of SUF4 and EC1 gene expression.


Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Fertilization/genetics , Gene Expression Regulation, Plant , Germ Cells, Plant/cytology , Germ Cells, Plant/metabolism , Trans-Activators/metabolism , Arabidopsis Proteins/metabolism , Conserved Sequence/genetics , Genes, Plant , Genes, Reporter , Green Fluorescent Proteins/metabolism , Nucleotide Motifs/genetics , Ovum/cytology , Ovum/metabolism , Phenotype , Promoter Regions, Genetic , Transcription, Genetic
12.
Plant Physiol ; 170(3): 1817-30, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26823545

ABSTRACT

Developmental or metabolic changes in chloroplasts can have profound effects on the rest of the plant cell. Such intracellular responses are associated with signals that originate in chloroplasts and convey information on their physiological status to the nucleus, which leads to large-scale changes in gene expression (retrograde signaling). A screen designed to identify components of retrograde signaling resulted in the discovery of the so-called genomes uncoupled (gun) mutants. Genetic evidence suggests that the chloroplast protein GUN1 integrates signals derived from perturbations in plastid redox state, plastid gene expression, and tetrapyrrole biosynthesis (TPB) in Arabidopsis (Arabidopsis thaliana) seedlings, exerting biogenic control of chloroplast functions. However, the molecular mechanism by which GUN1 integrates retrograde signaling in the chloroplast is unclear. Here we show that GUN1 also operates in adult plants, contributing to operational control of chloroplasts. The gun1 mutation genetically interacts with mutations of genes for the chloroplast ribosomal proteins S1 (PRPS1) and L11. Analysis of gun1 prps1 lines indicates that GUN1 controls PRPS1 accumulation at the protein level. The GUN1 protein physically interacts with proteins involved in chloroplast protein homeostasis based on coimmunoprecipitation experiments. Furthermore, yeast two-hybrid and bimolecular fluorescence complementation experiments suggest that GUN1 might transiently interact with several TPB enzymes, including Mg-chelatase subunit D (CHLD) and two other TPB enzymes known to activate retrograde signaling. Moreover, the association of PRPS1 and CHLD with protein complexes is modulated by GUN1. These findings allow us to speculate that retrograde signaling might involve GUN1-dependent formation of protein complexes.


Subject(s)
Arabidopsis Proteins/metabolism , Chloroplast Proteins/metabolism , DNA-Binding Proteins/metabolism , Homeostasis , Ribosomal Proteins/metabolism , Amino Acid Sequence , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Chloroplast Proteins/genetics , DNA-Binding Proteins/genetics , Epistasis, Genetic , Gene Expression Regulation, Plant , Immunoblotting , Lyases/genetics , Lyases/metabolism , Mutation , Plants, Genetically Modified , Plastids/genetics , Plastids/metabolism , Protein Binding , Protein Subunits/genetics , Protein Subunits/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Ribosomal Proteins/genetics , Sequence Homology, Amino Acid , Tetrapyrroles/biosynthesis
13.
Europace ; 19(6): 1002-1006, 2017 Jun 01.
Article in English | MEDLINE | ID: mdl-27353323

ABSTRACT

AIMS: Pocket haematoma is a common complication after defibrillator [implantable cardioverter defibrillator (ICD)] implantation, which is not only painful, but also increases the risk of device-related infection, and possibly embolic events. The present study seeks to evaluate the rate and predictors of clinically significant pocket haematoma. METHODS AND RESULTS: This study included 2500 patients receiving an ICD in the SIMPLE trial. A clinically significant pocket haematoma was defined as a haematoma that required re-operation or interruption of oral anticoagulation (OAC) therapy. Clinically significant pocket haematoma occurred in 56 of 2500 patients (2.2%) of which 6 (10.7%) developed device-related infection. Patients who developed pocket haematoma were older (mean age 67.6 ± 8.8 years vs. 62.7 ± 11.6 years, P < 0.001), were more likely to have permanent atrial fibrillation (30.4 vs. 6.7%, P < 0.001) and a history of stroke (17.9 vs. 6.7%, P = 0.004), or were more likely to receive peri-operative OAC (50.0 vs. 28.4%, P < 0.001), unfractionated heparin (16.1 vs. 5.2%, P = 0.003), or low-molecular-weight heparin (37.5 vs. 17.5%, P < 0.001). Independent predictors of wound haematoma on multivariable analysis included the use of heparin bridging (OR 2.65, 95% CI 1.48-4.73, P = 0.001), sub-pectoral location of ICD (OR 2.00, 95% CI 1.12-3.57, P =0.020), previous stroke (OR 2.47, 95% CI 1.20-5.10, P = 0.015), an upgrade from permanent pacemaker (OR 2.52, 95% CI 1.07-5.94, P = 0.035), and older age (OR 1.03, 95% CI 1.00-1.06, P = 0.049). CONCLUSION: Pocket haematoma remains an important complication of ICD implantation and is associated with a high risk of infection. Independent predictors of pocket haematoma include heparin bridging, prior stroke, sub-pectoral placement of ICD, older age, and upgrade from a pacemaker.


Subject(s)
Arrhythmias, Cardiac/therapy , Defibrillators, Implantable , Electric Countershock/instrumentation , Hematoma/epidemiology , Prosthesis Implantation/instrumentation , Surgical Wound/epidemiology , Age Factors , Aged , Arrhythmias, Cardiac/diagnosis , Arrhythmias, Cardiac/mortality , Chi-Square Distribution , Defibrillators, Implantable/adverse effects , Electric Countershock/adverse effects , Electric Countershock/mortality , Female , Hematoma/diagnosis , Heparin/administration & dosage , Heparin/adverse effects , Humans , Incidence , Logistic Models , Male , Middle Aged , Multivariate Analysis , Nonlinear Dynamics , Odds Ratio , Prospective Studies , Prosthesis Implantation/adverse effects , Prosthesis Implantation/mortality , Prosthesis-Related Infections/epidemiology , Risk Factors , Surgical Wound/diagnosis , Treatment Outcome
14.
Proc Natl Acad Sci U S A ; 111(41): 14947-52, 2014 Oct 14.
Article in English | MEDLINE | ID: mdl-25271326

ABSTRACT

To cope with growth in low-phosphate (Pi) soils, plants have evolved adaptive responses that involve both developmental and metabolic changes. Phosphate Starvation Response 1 (PHR1) and related transcription factors play a central role in the control of Pi starvation responses (PSRs). How Pi levels control PHR1 activity, and thus PSRs, remains to be elucidated. Here, we identify a direct Pi-dependent inhibitor of PHR1 in Arabidopsis, SPX1, a nuclear protein that shares the SPX domain with yeast Pi sensors and with several Pi starvation signaling proteins from plants. Double mutation of SPX1 and of a related gene, SPX2, resulted in molecular and physiological changes indicative of increased PHR1 activity in plants grown in Pi-sufficient conditions or after Pi refeeding of Pi-starved plants but had only a limited effect on PHR1 activity in Pi-starved plants. These data indicate that SPX1 and SPX2 have a cellular Pi-dependent inhibitory effect on PHR1. Coimmunoprecipitation assays showed that the SPX1/PHR1 interaction in planta is highly Pi-dependent. DNA-binding and pull-down assays with bacterially expressed, affinity-purified tagged SPX1 and ΔPHR1 proteins showed that SPX1 is a competitive inhibitor of PHR1 binding to its recognition sequence, and that its efficiency is highly dependent on the presence of Pi or phosphite, a nonmetabolizable Pi analog that can repress PSRs. The relative strength of the SPX1/PHR1 interaction is thus directly influenced by Pi, providing a link between Pi perception and signaling.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Nuclear Proteins/metabolism , Phosphates/pharmacology , Transcription Factors/metabolism , Arabidopsis/drug effects , Arabidopsis Proteins/antagonists & inhibitors , Arabidopsis Proteins/genetics , DNA, Plant/metabolism , Models, Biological , Mutation/genetics , Nuclear Proteins/genetics , Protein Binding/drug effects , Transcription Factors/antagonists & inhibitors
15.
Plant Cell ; 25(7): 2560-72, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23847151

ABSTRACT

MADS domain transcription factors are key regulators of eukaryotic development. In plants, the homeotic MIKC MADS factors that regulate floral organ identity have been studied in great detail. Based on genetic and protein-protein interaction studies, a floral quartet model was proposed that describes how these MADS domain proteins assemble into higher order complexes to regulate their target genes. However, despite the attractiveness of this model and its general acceptance in the literature, solid in vivo proof has never been provided. To gain deeper insight into the mechanisms of transcriptional regulation by MADS domain factors, we studied how SEEDSTICK (STK) and SEPALLATA3 (SEP3) directly regulate the expression of the reproductive meristem gene family transcription factor-encoding gene VERDANDI (VDD). Our data show that STK-SEP3 dimers can induce loop formation in the VDD promoter by binding to two nearby CC(A/T)6GG (CArG) boxes and that this is essential for promoter activity. Our in vivo data show that the size and position of this loop, determined by the choice of CArG element usage, is essential for correct expression. Our studies provide solid in vivo evidence for the floral quartet model.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , DNA, Plant/metabolism , Homeodomain Proteins/metabolism , MADS Domain Proteins/metabolism , Transcription Factors/metabolism , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis Proteins/genetics , Base Sequence , DNA, Plant/chemistry , DNA, Plant/genetics , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , Homeodomain Proteins/genetics , Immunoblotting , In Situ Hybridization , MADS Domain Proteins/genetics , Mutation , Nucleic Acid Conformation , Nucleotide Motifs/genetics , Ovule/genetics , Ovule/growth & development , Ovule/metabolism , Plants, Genetically Modified , Promoter Regions, Genetic/genetics , Protein Binding , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Nucleic Acid , Transcription Factors/genetics
16.
Evol Dev ; 17(1): 82-91, 2015.
Article in English | MEDLINE | ID: mdl-25627715

ABSTRACT

One successful mechanism of seed dispersal in plants involves production of edible fleshy structures which attract frugivorous animals and transfer this task to them. Not only Angiosperms but also Gymnosperms may use the fleshy fruit habit for seed dispersal, and a similar suite of MADS-box genes may be expressed as these structures form. Magnolia grandiflora produces dry follicles which, at maturity, open to reveal brightly colored fleshy seeds. This species thus also employs endozoochory for seed dispersal, although it produces dry fruits. Molecular analysis reveals that genes involved in softening and color changes are expressed at late stages of seed development, when the fleshy seed sarcotesta softens and accumulates carotenoids. Several MADS-box genes have also been studied and results highlight the existence of a basic genetic toolkit which may be common to all fleshy fruit-like structures, independently of their anatomic origin. According to their expression patterns, one of two AGAMOUS genes and the three SEPALLATA genes known so far in Magnolia are of particular interest. Duplication of AGAMOUS already occurs in both Nymphaeales and Magnoliids, although the lack of functional gene analysis prevents comparisons with known duplications in the AGAMOUS lineage of core Eudicots.


Subject(s)
Magnolia/genetics , Seeds/metabolism , Evolution, Molecular , Fruit/anatomy & histology , Fruit/chemistry , Gene Expression Regulation, Plant , MADS Domain Proteins/genetics , Magnolia/embryology , Magnolia/metabolism , Phylogeny , Plant Proteins/genetics , Real-Time Polymerase Chain Reaction , Seeds/anatomy & histology , Transcriptome
17.
J Integr Plant Biol ; 57(11): 892-901, 2015 Nov.
Article in English | MEDLINE | ID: mdl-25966787

ABSTRACT

In recent years, peptide aptamers have emerged as novel molecular tools that have attracted the attention of researchers in various fields of basic and applied science, ranging from medicine to analytical chemistry. These artificial short peptides are able to specifically bind, track, and inhibit a given target molecule with high affinity, even molecules with poor immunogenicity or high toxicity, and represent a remarkable alternative to antibodies in many different applications. Their use is on the rise, driven mainly by the medical and pharmaceutical sector. Here we discuss the enormous potential of peptide aptamers in both basic and applied aspects of plant biotechnology and food safety. The different peptide aptamer selection methods available both in vivo and in vitro are introduced, and the most important possible applications in plant biotechnology are illustrated. In particular, we discuss the generation of broad-based virus resistance in crops, "reverse genetics" and aptasensors in bioassays for detecting contaminations in food and feed. Furthermore, we suggest an alternative to the transfer of peptide aptamers into plant cells via genetic transformation, based on the use of cell-penetrating peptides that overcome the limits imposed by both crop transformation and Genetically Modified Organism commercialization.


Subject(s)
Aptamers, Peptide , Biotechnology/methods , Plant Proteins/antagonists & inhibitors , Biotechnology/trends , Food Safety , Genomics , Plant Immunity , Plant Viruses/immunology , Plants, Genetically Modified
18.
Plant J ; 76(3): 446-55, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23941199

ABSTRACT

Upon hormonal signaling, ovules develop as lateral organs from the placenta. Ovule numbers ultimately determine the number of seeds that develop, and thereby contribute to the final seed yield in crop plants. We demonstrate here that CUP-SHAPED COTYLEDON 1 (CUC1), CUC2 and AINTEGUMENTA (ANT) have additive effects on ovule primordia formation. We show that expression of the CUC1 and CUC2 genes is required to redundantly regulate expression of PINFORMED1 (PIN1), which in turn is required for ovule primordia formation. Furthermore, our results suggest that the auxin response factor MONOPTEROS (MP/ARF5) may directly bind ANT, CUC1 and CUC2 and promote their transcription. Based on our findings, we propose an integrative model to describe the molecular mechanisms of the early stages of ovule development.


Subject(s)
Arabidopsis Proteins/physiology , Arabidopsis/embryology , Ovule/embryology , Transcription Factors/physiology , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , DNA-Binding Proteins/metabolism , Green Fluorescent Proteins , Membrane Transport Proteins/metabolism , Models, Biological , Promoter Regions, Genetic , Transcription Factors/metabolism
19.
BMC Plant Biol ; 14: 319, 2014 Nov 30.
Article in English | MEDLINE | ID: mdl-25433802

ABSTRACT

BACKGROUND: The identity of flower organs is specified by various MIKC MADS-box transcription factors which act in a combinatorial manner. TM8 is a MADS-box gene that was isolated from the floral meristem of a tomato mutant more than twenty years ago, but is still poorly known from a functional point of view in spite of being present in both Angiosperms and Gymnosperms, with some species harbouring more than one copy of the gene. This study reports a characterization of TM8 that was carried out in transgenic tomato plants with altered expression of the gene. RESULTS: Tomato plants over-expressing either TM8 or a chimeric repressor form of the gene (TM8:SRDX) were prepared. In the TM8 up-regulated plants it was possible to observe anomalous stamens with poorly viable pollen and altered expression of several floral identity genes, among them B-, C- and E-function ones, while no apparent morphological modifications were visible in the other whorls. Oblong ovaries and fruits, that were also parthenocarpic, were obtained in the plants expressing the TM8:SRDX repressor gene. Such ovaries showed modified expression of various carpel-related genes. No apparent modifications could be seen in the other flower whorls. The latter plants had also epinastic leaves and malformed flower abscission zones. By using yeast two hybrid assays it was possible to show that TM8 was able to interact in yeast with MACROCALIX. CONCLUSIONS: The impact of the ectopically altered TM8 expression on the reproductive structures suggests that this gene plays some role in the development of the tomato flower. MACROCALYX, a putative A-function MADS-box gene, was expressed in all the four whorls of fully developed flowers, and showed quantitative variations that were opposite to those of TM8 in the anomalous stamens and ovaries. Since the TM8 protein interacted in vitro only with the A-function MADS-box protein MACROCALYX, it seems that for the correct differentiation of the tomato reproductive structures possible interactions between TM8 and MACROCALYX proteins might be important.


Subject(s)
Gene Expression Regulation, Plant , MADS Domain Proteins/genetics , Plant Proteins/genetics , Solanum lycopersicum/genetics , Flowers/genetics , Flowers/growth & development , Flowers/metabolism , Gene Expression Regulation, Developmental , Solanum lycopersicum/growth & development , Solanum lycopersicum/metabolism , MADS Domain Proteins/metabolism , Molecular Sequence Data , Plant Proteins/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/growth & development , Plants, Genetically Modified/metabolism , Real-Time Polymerase Chain Reaction , Sequence Analysis, DNA
20.
J Exp Bot ; 65(18): 5459-71, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25053647

ABSTRACT

Arabinogalactan proteins (AGPs) are heavily glycosylated proteins existing in all members of the plant kingdom and are differentially distributed through distinctive developmental stages. Here, we showed the individual distributions of specific Arabidopsis AGPs: AGP1, AGP9, AGP12, AGP15, and AGP23, throughout reproductive tissues and indicated their possible roles in several reproductive processes. AGP genes specifically expressed in female tissues were identified using available microarray data. This selection was confirmed by promoter analysis using multiple green fluorescent protein fusions to a nuclear localization signal, ß-glucuronidase fusions, and in situ hybridization as approaches to confirm the expression patterns of the AGPs. Promoter analysis allowed the detection of a specific and differential presence of these proteins along the pathway followed by the pollen tube during its journey to reach the egg and the central cell inside the embryo sac. AGP1 was expressed in the stigma, style, transmitting tract, and the chalazal and funiculus tissues of the ovules. AGP9 was present along the vasculature of the reproductive tissues and AGP12 was expressed in the stigmatic cells, chalazal and funiculus cells of the ovules, and in the septum. AGP15 was expressed in all pistil tissues, except in the transmitting tract, while AGP23 was specific to the pollen grain and pollen tube. The expression pattern of these AGPs provides new evidence for the detection of a subset of specific AGPs involved in plant reproductive processes, being of significance for this field of study. AGPs are prominent candidates for male-female communication during reproduction.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Flowers/metabolism , Mucoproteins/metabolism , Pollen Tube/metabolism , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Mucoproteins/genetics , Plant Proteins/genetics , Plant Proteins/metabolism
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