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1.
Sci Total Environ ; 803: 150006, 2022 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-34487902

RESUMO

Soil contamination with trace metal(loid) elements (TME) is a global concern. This has focused interest on TME-tolerant plants, some of which can hyperaccumulate extraordinary amounts of TME into above-ground tissues, for potential treatment of these soils. However, intra-species variability in TME hyperaccumulation is not yet sufficiently understood to fully harness this potential. Particularly, little is known about the rhizosphere microbial communities associated with hyperaccumulating plants and whether or not they facilitate TME uptake. The aim of this study is to characterize the diversity and structure of Arabidopsis halleri rhizosphere-influenced and background (i.e., non-Arabidopsis) soil microbial communities in four plant populations with contrasting Zn and Cd hyperaccumulation traits, two each from contaminated and uncontaminated sites. Microbial community properties were assessed along with geographic location, climate, abiotic soil properties, and plant parameters to explain variation in Zn and Cd hyperaccumulation. Site type (TME-contaminated vs. uncontaminated) and location explained 44% of bacterial/archaeal and 28% of fungal community variability. A linear discriminant effect size (LEfSe) analysis identified a greater number of taxa defining rhizosphere microbial communities than associated background soils. Further, in TME-contaminated soils, the number of rhizosphere-defining taxa was 6-fold greater than in the background soils. In contrast, the corresponding ratio for uncontaminated sites, was 3 and 1.6 for bacteria/archaea and fungi, respectively. The variables analyzed explained 71% and 76% of the variance in Zn and Cd hyperaccumulation, respectively; however, each hyperaccumulation pattern was associated with different variables. A. halleri rhizosphere fungal richness and diversity associated most strongly with Zn hyperaccumulation, whereas soil Cd and Zn bioavailability had the strongest associations with Cd hyperaccumulation. Our results indicate strong associations between A. halleri TME hyperaccumulation and rhizosphere microbial community properties, a finding that needs to be further explored to optimize phytoremediation technology that is based on hyperaccumulation.


Assuntos
Arabidopsis , Microbiota , Poluentes do Solo , Biodegradação Ambiental , Cádmio , Rizosfera , Solo , Microbiologia do Solo , Poluentes do Solo/análise , Zinco
2.
Chemosphere ; 286(Pt 1): 131633, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34325267

RESUMO

Endocrine-disrupting chemicals (EDCs) are widespread contaminants that severely affect the endocrine systems of living organisms. In addition to the conventional instrument-based approaches for quantifying organic pollutants, a monitoring method using transgenic plants has also been proposed. Plants carrying a recombinant receptor gene combined with a reporter gene represent a system for the easy detection of ligands that specifically bind to the receptor molecule. Here, the EDC detection sensitivity of transgenic Arabidopsis plants expressing the medaka (Oryzias latipes) estrogen receptor (mER) and green fluorescent protein (GFP) genes, was assessed. Four transgenic Arabidopsis lines, obtained by transformation with expression plasmids constructed using combinations of two types of the ligand-binding domains of mER, the DNA-binding domain of LexA and the transactivation domain of VP16 in the chimeric receptors, showed significant induction of GFP when germinated on a medium contaminated with 1 ng/mL 4-t-octylphenol (OP). The most sensitive XmEV19-2 plants detected 0.1 ng/mL OP and 1 pg/mL 17ß-estradiol. GFP expression was suppressed by the insecticides imidacloprid and fipronil, whereas perfluorooctanesulfonic acid induced it at 0.1 ng/mL. Experiments with river water-based medium showed that XmEV19-2 can be used for monitoring polluted waters, detecting OP at concentrations as low as 5 ng/mL. Notably, XmEV19-2 showed a significant decrease in root length when grown on 0.1 ng/mL OP. mER transgenic plants can be a promising tool for simple monitoring of EDCs, without the need for extraction and concentration steps in sample preparation.


Assuntos
Arabidopsis , Disruptores Endócrinos , Oryzias , Poluentes Químicos da Água , Animais , Arabidopsis/genética , Disruptores Endócrinos/análise , Disruptores Endócrinos/toxicidade , Monitoramento Ambiental , Oryzias/genética , Plantas Geneticamente Modificadas/genética , Receptores de Estrogênio/genética , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidade
3.
Environ Pollut ; 292(Pt B): 118448, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34728324

RESUMO

Application of nanopesticides may substantially increase surface attachment and internalization of engineered nanoparticles (ENPs) in food crops. This study investigated the role of stomata in the internalization of silver nanoparticles (Ag NPs) using abscisic acid (ABA)-responsive ecotypes (Ler and Col-7) and ABA-insensitive mutants (ost1-2 and scord7) of Arabidopsis thaliana in batch sorption experiments, in combination with microscopic visualization. Compared with those of the ABA-free control, stomatal apertures were significantly smaller for the Ler and Col-7 ecotypes (p ˂ 0.05) but remained unchanged for the ost1-2 and scord7 mutants, after exposure to 10 µM ABA for 1 h. Generally Ag NP sorption to the leaves of the Ler and Col-7 ecotypes treated with 10 µM ABA was lower than that in the ABA-free control, mainly due to ABA-induced stomatal closure. The difference in Ag NP sorption with and without ABA was less pronounced for Col-7 than for Ler, suggesting different sorption behaviors between these two ecotypes. In contrast, there was no significant difference in foliar sorption of Ag NPs by the ost1-2 and scord7 mutants with and without ABA treatment. Ag NPs were widely attached to the Arabidopsis leaf surface, and found at cell membrane, cytoplasm, and plasmodesmata, as revealed by scanning electron microscopy and transmission electron microscopy, respectively. These results highlight the important role of stomata in the internationalization of ENPs in plants and may have broad implications in foliar application of nanopesticides and minimizing contamination of food crops by ENPs.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Nanopartículas Metálicas , Ácido Abscísico , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Mutação , Estômatos de Plantas/metabolismo , Proteínas Quinases/metabolismo , Espécies Reativas de Oxigênio , Transdução de Sinais , Prata
4.
Sci Total Environ ; 805: 150335, 2022 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-34818777

RESUMO

Organic acids play an important role in metal tolerance, uptake, and translocation in hyperaccumulators. Phytolacca americana is a rare earth element (REE) hyperaccumulator, but the underlying mechanisms on REE tolerance and accumulation mediated by organic acids are poorly understood. Here, we reported for the first time the strategy of P. americana to enhance REE tolerance and accumulation through organic acids from root external secretion to internal biosynthesis. Different from the exclusion of heavy metal by organic acid in the typical plants, the results showed that oxalate secretion (0.3-0.6 µmol h-1 g-1 root DW) induced by yttrium (Y) could not prevent Y from entering the roots, resulting in excess Y uptake by P. americana. Yttrium stress also stimulated the accumulation of malate and citrate by 1.4- and 2.0-folds in the root cortex. Exogenous malate and citrate promoted the redistribution of Y from the root cell walls to the shoot by 30% and 21%, respectively. Based on comparative transcriptome analysis, 6-fold up-regulation was observed in PaNIP1;2, whose homology AtNIP1;2 is responsible for the transport of Al-malate in Arabidopsis. These results suggested that the promoted formation of Y-malate complexes within the roots potentially accelerated the transport of Y from P. americana roots to shoots through PaNIP1;2. Our study revealed the potential mechanism of organic acids in the external exclusion and internal detoxification and translocation of REE in P. americana roots, which provided a basis for improving the efficiency of REE phytoextraction.


Assuntos
Arabidopsis , Metais Terras Raras , Phytolacca americana , Compostos Orgânicos , Raízes de Plantas
5.
BMC Genomics ; 22(1): 817, 2021 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-34772363

RESUMO

BACKGROUND: S-domain receptor-like kinases (SD-RLKs) are an important and multi-gene subfamily of plant receptor-like/pelle kinases (RLKs), which are known to play a significant role in the development and immune responses of Arabidopsis thaliana. The conserved cysteine residues in the extracellular domain of SD-RLKs make them interesting candidates for sensing reactive oxygen species (ROS), assisting oxidative stress mitigation and associated signaling pathways during abiotic stresses. However, how closely SD-RLKs are interrelated to abiotic stress mitigation and signaling remains unknown in A. thaliana. RESULTS: This study was initiated by examining the chromosomal localization, phylogeny, sequence and differential expression analyses of 37 SD-RLK genes using publicly accessible microarray datasets under cold, osmotic stress, genotoxic stress, drought, salt, UV-B, heat and wounding. Out of 37 SD-RLKs, 12 genes displayed differential expression patterns in both the root and the shoot tissues. Promoter structure analysis suggested that these 12 SD-RLK genes harbour several potential cis-regulatory elements (CREs), which are involved in regulating multiple abiotic stress responses. Based on these observations, we investigated the expression patterns of 12 selected SD-RLKs under ozone, wounding, oxidative (methyl viologen), UV-B, cold, and light stress at different time points using semi-qRT-PCR. Of these 12 SD-SRKs, the genes At1g61360, At1g61460, At1g61380, and At4g27300 emerged as potential candidates that maintain their expression in most of the stress treatments till exposure for 12 h. Expression patterns of these four genes were further verified under similar stress treatments using qRT-PCR. The expression analysis indicated that the gene At1g61360, At1g61380, and At1g61460 were mostly up-regulated, whereas the expression of At4g27300 either up- or down-regulated in these conditions. CONCLUSIONS: To summarize, the computational analysis and differential transcript accumulation of SD-RLKs under various abiotic stresses suggested their association with abiotic stress tolerance and related signaling in A. thaliana. We believe that a further detailed study will decipher the specific role of these representative SD-RLKs in abiotic stress mitigation vis-a-vis signaling pathways in A. thaliana.


Assuntos
Arabidopsis , Arabidopsis/genética , Simulação por Computador , Secas , Regulação da Expressão Gênica de Plantas , Filogenia , Cloreto de Sódio , Estresse Fisiológico/genética
6.
J Biotechnol ; 342: 102-113, 2021 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-34736953

RESUMO

Indole alkaloid camalexin has potential medicinal properties such as suppressing the viability of leukemic but not normal cells. Camalexin is not produced in plants and an external factor is required to activate its biosynthesis. In this work, we stimulated camalexin biosynthesis in Arabidopsis calli by blocking one of repressors of the jasmonate pathway, the jasmonate ZIM-domain protein 1 (JAZ1) by using amiRNA targeting JAZ1 gene transcripts. Inhibition of the JAZ1 gene led to an increase in camalexin content from trace amounts in control culture to 9 µg/g DW in the jaz1 line without affecting growth. In addition, JAZ1 silencing enhanced tolerance to cold stress with simultaneous increasing camalexin content up to 30 µg/g DW. Real-time quantitative PCR determination of marker gene expression showed that effects caused by the JAZ1 silencing might be realized through crosslinking JA, ROS, and abscisic acid signaling pathways. Thus, targeting the distal components of signaling pathways can be suggested as a tool for bioengineering of secondary metabolism, along with standard techniques for targeting biosynthetic genes or genes encoding transcription factors.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Ciclopentanos , Regulação da Expressão Gênica de Plantas , Indóis , Oxilipinas , Proteínas Repressoras/metabolismo , Metabolismo Secundário , Tiazóis
7.
Planta ; 255(1): 5, 2021 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-34841457

RESUMO

MAIN CONCLUSION: Plant CLE peptides, which regulate stem cell maintenance in shoot and root meristems and in vascular bundles through LRR family receptor kinases, are novel, complex, and to some extent conserved. Over the past two decades, peptide ligands of the CLAVATA3 (CLV3) /Embryo Surrounding Region (CLE) family have been recognized as critical short- and long-distance communication signals in plants, especially for stem cell homeostasis, cell fate determination and physiological responses. Stem cells located at the shoot apical meristem (SAM), the root apical meristem (RAM) and the procambium divide and differentiate into specialized cells that form a variety of tissues such as epidermis, ground tissues, xylem and phloem. In the SAM of Arabidopsis (Arabidopsis thaliana), the CLV3 peptide restricts the number of stem cells via leucine-rich repeat (LRR)-type receptor kinases. In the RAM, root-active CLE peptides are critical negative regulators, while ROOT GROWTH FACTOR (RGF) peptides are positive regulators in stem cell maintenance. Among those root-active CLE peptides, CLE25 promotes, while CLE45 inhibits phloem differentiation. In vascular bundles, TRACHEARY ELEMENT DIFFERENTIATION INHIBITORY FACTOR (TDIF)/CLE41/CLE44 promotes procambium cell division, and prevents xylem differentiation. Orthologs of CLV3 have been identified in liverwort (Marchantia polymorpha), tomato (Solanum lycopersicum), rice (Oryza sativa), maize (Zea mays) and lotus (Lotus japonicas), suggesting that CLV3 is an evolutionarily conserved signal in stem cell maintenance. However, functional characterization of endogenous CLE peptides and corresponding receptor kinases, and the downstream signal transduction has been challenging due to their genome-wide redundancies and rapid evolution.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Membrana , Meristema/metabolismo , Peptídeos/metabolismo , Células-Tronco/metabolismo
8.
Planta ; 254(6): 131, 2021 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-34821996

RESUMO

MAIN CONCLUSION: Multiple rice GT61 members were demonstrated to be xylan arabinosyltransferases (XATs) mediating 3-O-arabinosylation of xylan and the functions of XATs and xylan 2-O-xylosyltransferases were shown to be conserved in grass species. Xylan is the major hemicellulose in the cell walls of grass species and it is typified by having arabinofuranosyl (Araf) substitutions. In this report, we demonstrated that four previously uncharacterized, Golgi-localized glycosyltransferases residing in clade A or B of the rice GT61 family were able to mediate 3-O-arabinosylation of xylan when heterologously expressed in the Arabidopsis gux1/2/3 triple mutant. Biochemical characterization of their recombinant proteins established that they were xylan arabinosyltransferases (XATs) capable of transferring Araf residues onto xylohexaose acceptors, and thus they were named OsXAT4, OsXAT5, OsXAT6 and OsXAT7. OsXAT5 and the previously identified OsXAT2 were shown to be able to arabinosylate xylooligomers with a degree of polymerization of as low as 3. Furthermore, a number of XAT homologs from maize, sorghum, Brachypodium and switchgrass were found to exhibit activities catalyzing Araf transfer onto xylohexaose, indicating that they are XATs involved in xylan arabinosylation in these grass species. Moreover, we revealed that homologs of another GT61 member, xylan 2-O-xylosyltransferase (XYXT1), from these grass species could mediate 2-O-xylosylation of xylan when expressed in the Arabidopsis gux1/2/3 mutant. Together, our findings indicate that multiple OsXATs are involved in 3-O-arabinosylation of xylan and the functions of XATs and XYXTs are conserved in grass species.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Parede Celular , Glicosiltransferases/genética , Xilanos
9.
Plant Sci ; 313: 111047, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34763851

RESUMO

The dehydration-responsive element-binding (DREB) transcription factors play important roles in regulation of plant responses to abiotic stresses, however, few DREBs have been isolated from a desiccation tolerance moss, and the role of DREBs in the DT mechanism is still unknown. We have functionally characterized a unique DREB transcription factor BaDBL1 from the DT moss Bryum argenteum. Expression pattern analysis revealed that BaDBL1 was induced by dehydration-rehydration, salt, cold, and abscisic acid treatments. BaDBL1 was localized in the nucleus and had a transactivation region in its C-terminal region. Overexpression of BaDBL1 in Arabidopsis resulted in significantly increased osmotic and salt stress tolerance, as illustrated by higher fresh weight and antioxidase activities (SOD, POD and CAT) compared with WT under osmotic and salt stresses. Moreover, the transcription of stress-responsive genes, such as AtRD29A and AtCOR15A, AtLEA in BaDBL1-overexpressing lines were significantly up-regulated under osmotic and salt stresses compared with WT. Transcriptomic analysis revealed that BaDBL1-overexpression affected the lignin biosynthesis pathway by improving lignin content and regulating lignin-biosynthesis-related genes under osmotic stress. The results suggest that BaDBL1 may regulate plant tolerance to stress by enhancing anti-oxidase activities, regulating expression of stress-related genes and effecting the lignin biosynthesis, making BaDBL1 a candidate gene for stress tolerance improvement in crops.


Assuntos
Arabidopsis/genética , Bryopsida/genética , Desidratação/genética , Secas , Pressão Osmótica/fisiologia , Tolerância ao Sal/genética , Fatores de Transcrição/genética , Arabidopsis/fisiologia , Bryopsida/fisiologia , Desidratação/fisiopatologia , Regulação da Expressão Gênica de Plantas , Técnicas de Transferência de Genes , Genes de Plantas , Filogenia , Plantas Geneticamente Modificadas , Tolerância ao Sal/fisiologia
10.
Plant Sci ; 313: 111073, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34763865

RESUMO

MicroRNAs (miRNAs), a class of single-stranded non-coding RNA of 20-24 nucleotides, regulate gene expression by target gene transcript cleavage or translation inhibition. The phytohormone auxin is a crucial regulator of almost every process involved in plant growth and development. Several studies have demonstrated the involvement of miRNA(s) in the regulation of the auxin signaling pathway and plant development. However, very few studies have identified the auxin-mediated regulation of miRNA(s). In this study, we reveal the detailed mechanism of auxin-mediated regulation of the cell wall-related miR775- Galactosyl transferase (GalT) module, which plays an important role in root growth in Arabidopsis thaliana. We also showed two interdependent mechanisms by which miR775 regulates root growth: miR775-GalT and light-mediated sucrose-dependent pathways. Treatment of GUS reporter lines with Indole Acetic Acid (IAA), sucrose, and light apparently enhanced the abundance of miR775 in root tissue. miR775 overexpressing (miR775OX) lines showed changes in root architecture, including increased primary root growth and root hair, by targeting GalT. miR775OX lines also showed tolerance toward low Pi. These results provide new insights into the auxin regulation of cell wall-related miR775 and suggest its significant role in plant root growth and development by modifying the cell wall.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Ácidos Indolacéticos/metabolismo , MicroRNAs/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/genética , Sacarose/metabolismo , Adaptação Ocular/efeitos dos fármacos , Adaptação Ocular/genética , Regulação da Expressão Gênica de Plantas , Variação Genética , Genótipo , Crescimento e Desenvolvimento/genética , Redes e Vias Metabólicas/genética
11.
Plant Sci ; 313: 111086, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34763871

RESUMO

Plants are sessile and lack behavioural responses to avoid extreme environmental changes linked to annual seasons. For survival, they have evolved elaborate sensory systems coordinating their architecture and physiology with fluctuating diurnal and seasonal temperatures. PHYTOCHROME-INTERACTING FACTOR 4 (PIF4) was initially identified as a key component of the Arabidopsis thaliana phytochrome signalling pathway. It was then identified as playing a central role in promoting plant hypocotyl growth via the activation of auxin synthesis and signalling-related genes. Recent studies expanded its known regulatory functions to thermomorphogenesis and defined PIF4 as a central molecular hub for the integration of environmental light and temperature cues. The present review comprehensively summarizes recent progress in our understanding of PIF4 function in Arabidopsis thaliana, including PIF4-mediated photomorphogenesis and thermomorphogenesis, and the contribution of PIF4 to plant growth via the integration of environmental light and temperature cues. Remaining questions and possible directions for future research on PIF4 are also discussed.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Hipocótilo/crescimento & desenvolvimento , Hipocótilo/genética , Fitocromo/metabolismo , Desenvolvimento Vegetal/genética , Transdução de Sinais/genética , Transdução de Sinais/fisiologia , Adaptação Ocular/fisiologia , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Fitocromo/genética , Temperatura , Sensação Térmica/fisiologia
13.
Plant Mol Biol ; 107(3): 159-175, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34599731

RESUMO

KEY MESSAGE: A stress induced calcium-binding protein, RD20/CLO3 interacts with the alpha subunit of the heterotrimeric G-protein complex in Arabidopsis and affects etiolation and leaf morphology. Heterotrimeric G proteins and calcium signaling have both been shown to play a role in the response to environmental abiotic stress in plants; however, the interaction between calcium-binding proteins and G-protein signaling molecules remains elusive. We investigated the interaction between the alpha subunit of the heterotrimeric G-protein complex, GPA1, of Arabidopsis thaliana with the calcium-binding protein, the caleosin RD20/CLO3, a gene strongly induced by drought, salt and abscisic acid. The proteins were found to interact in vivo by bimolecular fluorescent complementation (BiFC); the interaction was localized to the endoplasmic reticulum and to oil bodies within the cell. The constitutively GTP-bound GPA1 (GPA1QL) also interacts with RD20/CLO3 as well as its EF-hand mutant variations and these interactions are localized to the plasma membrane. The N-terminal portion of RD20/CLO3 was found to be responsible for the interaction with GPA1 and GPA1QL using both BiFC and yeast two-hybrid assays. RD20/CLO3 contains a single calcium-binding EF-hand in the N-terminal portion of the protein; disruption of the calcium-binding capacity of the protein obliterates interaction with GPA1 in in vivo assays and decreases the interaction between the caleosin and the constitutively active GPA1QL. Analysis of rd20/clo3 mutants shows that RD20/CLO3 plays a key role in the signaling pathway controlling hypocotyl length in dark grown seedlings and in leaf morphology. Our findings indicate a novel role for RD20/CLO3 as a negative regulator of GPA1.


Assuntos
Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Proteínas de Ligação ao Cálcio/metabolismo , Subunidades alfa de Proteínas de Ligação ao GTP/genética , Estresse Fisiológico/fisiologia , Proteínas de Ligação ao Cálcio/genética , Escuridão , Subunidades alfa de Proteínas de Ligação ao GTP/metabolismo , Regulação da Expressão Gênica de Plantas , Hipocótilo/genética , Hipocótilo/crescimento & desenvolvimento , Mutação , Folhas de Planta/fisiologia , Plantas Geneticamente Modificadas , Tabaco/genética , Tabaco/metabolismo , Técnicas do Sistema de Duplo-Híbrido
14.
Nat Plants ; 7(10): 1379-1388, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34650259

RESUMO

Plants seem to take up exogenous RNA that was artificially designed to target specific genes, followed by activation of the RNA interference (RNAi) machinery. It is, however, not known whether plants use RNAs themselves as signalling molecules in plant-to-plant communication, other than evidence that an exchange of small RNAs occurs between parasitic plants and their hosts. Exogenous RNAs from the environment, if taken up by some living organisms, can indeed induce RNAi. This phenomenon has been observed in nematodes and insects, and host Arabidopsis cells secrete exosome-like extracellular vesicles to deliver plant small RNAs into Botrytis cinerea. Here we show that micro-RNAs (miRNAs) produced by plants act as signalling molecules affecting gene expression in other, nearby plants. Exogenous miRNAs, such as miR156 and miR399, trigger RNAi via a mechanism requiring both AGO1 and RDR6. This emphasizes that the production of secondary small interfering RNAs is required. This evidence highlights the existence of a mechanism in which miRNAs represent signalling molecules that enable communication between plants.


Assuntos
Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , MicroRNAs/genética , Interferência de RNA , RNA de Plantas/genética , Arabidopsis/metabolismo
15.
Nat Commun ; 12(1): 5969, 2021 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-34645811

RESUMO

The Yersinia outer protein J (YopJ) family effectors are widely deployed through the type III secretion system by both plant and animal pathogens. As non-canonical acetyltransferases, the enzymatic activities of YopJ family effectors are allosterically activated by the eukaryote-specific ligand inositol hexaphosphate (InsP6). However, the underpinning molecular mechanism remains undefined. Here we present the crystal structure of apo-PopP2, a YopJ family member secreted by the plant pathogen Ralstonia solanacearum. Structural comparison of apo-PopP2 with the InsP6-bound PopP2 reveals a substantial conformational readjustment centered in the substrate-binding site. Combining biochemical and computational analyses, we further identify a mechanism by which the association of InsP6 with PopP2 induces an α-helix-to-ß-strand transition in the catalytic core, resulting in stabilization of the substrate recognition helix in the target protein binding site. Together, our study uncovers the molecular basis governing InsP6-mediated allosteric regulation of YopJ family acetyltransferases and further expands the paradigm of fold-switching proteins.


Assuntos
Acetiltransferases/química , Apoproteínas/química , Arabidopsis/microbiologia , Proteínas de Bactérias/química , Ácido Fítico/química , Ralstonia solanacearum/química , Acetiltransferases/genética , Acetiltransferases/metabolismo , Regulação Alostérica , Apoproteínas/genética , Apoproteínas/metabolismo , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Domínio Catalítico , Clonagem Molecular , Cristalografia por Raios X , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Modelos Moleculares , Ácido Fítico/metabolismo , Ligação Proteica , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Domínios e Motivos de Interação entre Proteínas , Ralstonia solanacearum/enzimologia , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Especificidade por Substrato , Tabaco/microbiologia
16.
Nat Commun ; 12(1): 5976, 2021 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-34645826

RESUMO

In plants, cytosine DNA methylations (5mCs) can happen in three sequence contexts as CpG, CHG, and CHH (where H = A, C, or T), which play different roles in the regulation of biological processes. Although long Nanopore reads are advantageous in the detection of 5mCs comparing to short-read bisulfite sequencing, existing methods can only detect 5mCs in the CpG context, which limits their application in plants. Here, we develop DeepSignal-plant, a deep learning tool to detect genome-wide 5mCs of all three contexts in plants from Nanopore reads. We sequence Arabidopsis thaliana and Oryza sativa using both Nanopore and bisulfite sequencing. We develop a denoising process for training models, which enables DeepSignal-plant to achieve high correlations with bisulfite sequencing for 5mC detection in all three contexts. Furthermore, DeepSignal-plant can profile more 5mC sites, which will help to provide a more complete understanding of epigenetic mechanisms of different biological processes.


Assuntos
Arabidopsis/genética , Citosina/metabolismo , DNA de Plantas/genética , Epigênese Genética , Genoma de Planta , Oryza/genética , Arabidopsis/metabolismo , Ilhas de CpG , Metilação de DNA , DNA de Plantas/metabolismo , Aprendizado Profundo , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Nanoporos , Oryza/metabolismo , Análise de Sequência de DNA , Sulfitos/química
17.
Int J Mol Sci ; 22(19)2021 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-34638657

RESUMO

Phytohormone ABA regulates the expression of numerous genes to significantly affect seed dormancy, seed germination and early seedling responses to biotic and abiotic stresses. However, the function of many ABA-responsive genes remains largely unknown. In order to improve the ABA-related signaling network, we conducted a large-scale ABA phenotype screening. LSH, an important transcription factor family, extensively participates in seedling development and floral organogenesis in plants, but whether its family genes are involved in the ABA signaling pathway has not been reported. Here we describe a new function of the transcription factor LSH8 in an ABA signaling pathway. In this study, we found that LSH8 was localized in the nucleus, and the expression level of LSH8 was significantly induced by exogenous ABA at the transcription level and protein level. Meanwhile, seed germination and root length measurements revealed that lsh8 mutant lines were ABA insensitive, whereas LSH8 overexpression lines showed an ABA-hypersensitive phenotype. With further TMT labeling quantitative proteomic analysis, we found that under ABA treatment, ABA-responsive proteins (ARPs) in the lsh8 mutant presented different changing patterns with those in wild-type Col4. Additionally, the number of ARPs contained in the lsh8 mutant was 397, six times the number in wild-type Col4. In addition, qPCR analysis found that under ABA treatment, LSH8 positively mediated the expression of downstream ABA-related genes of ABI3, ABI5, RD29B and RAB18. These results indicate that in Arabidopsis, LSH8 is a novel ABA regulator that could specifically change the expression pattern of APRs to positively mediate ABA responses.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Transdução de Sinais/fisiologia , Fatores de Transcrição/metabolismo , Regulação da Expressão Gênica de Plantas/fisiologia , Germinação/fisiologia , Fenótipo , Proteômica/métodos , Sementes/metabolismo
18.
Int J Mol Sci ; 22(19)2021 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-34638672

RESUMO

Flowering is an important developmental process from vegetative to reproductive growth in plant; thus, it is necessary to analyze the genes involved in the regulation of flowering time. The MADS-box transcription factor family exists widely in plants and plays an important role in the regulation of flowering time. However, the molecular mechanism of GmFULc involved in the regulation of plant flowering is not very clear. In this study, GmFULc protein had a typical MADS domain and it was a member of MADS-box transcription factor family. The expression analysis revealed that GmFULc was induced by short days (SD) and regulated by the circadian clock. Compared to wild type (WT), overexpression of GmFULc in transgenic Arabidopsis caused significantly earlier flowering time, while ful mutants flowered later, and overexpression of GmFULc rescued the late-flowering phenotype of ful mutants. ChIP-seq of GmFULc binding sites identified potential direct targets, including TOPLESS (TPL), and it inhibited the transcriptional activity of TPL. In addition, the transcription levels of FLOWERING LOCUS T (FT), SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1) and LEAFY (LFY) in the downstream of TPL were increased in GmFULc- overexpressionArabidopsis, suggesting that the early flowering phenotype was associated with up-regulation of these genes. Our results suggested that GmFULc inhibited the transcriptional activity of TPL and induced expression of FT, SOC1 and LFY to promote flowering.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Plantas Geneticamente Modificadas/genética , Soja/genética , Relógios Circadianos/genética , Flores/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Regulação da Expressão Gênica de Plantas/genética , Fotoperíodo , Folhas de Planta/genética , Reprodução/genética
19.
Int J Mol Sci ; 22(19)2021 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-34638681

RESUMO

Reactive oxygen species (ROS) signalling is crucial in modulating stress responses in plants, and NADPH oxidases (NOXs) are an important component of signal transduction under salt stress. The goal of this research was to investigate whether the regulation of NOX-dependent signalling during mild and severe salinity differs between the halophyte Eutrema salsugineum and the glycophyte Arabidopsis thaliana. Gene expression analyses showed that salt-induced expression patterns of two NOX genes, RBOHD and RBOHF, varied between the halophyte and the glycophyte. Five days of salinity stimulated the expression of both genes in E. salsugineum leaves, while their expression in A. thaliana decreased. This was not accompanied by changes in the total NOX activity in E. salsugineum, while the activity in A. thaliana was reduced. The expression of the RBOHD and RBOHF genes in E. salsugineum leaves was induced by abscisic acid (ABA) and ethephon spraying. The in silico analyses of promoter sequences of RBOHD and RBOHF revealed multiple cis-acting elements related to hormone responses, and their distribution varied between E. salsugineum and A. thaliana. Our results indicate that, in the halophyte E. salsugineum, the maintenance of the basal activity of NOXs in leaves plays a role during acclimation responses to salt stress. The different expression patterns of the RBOHD and RBOHF genes under salinity in E. salsugineum and A. thaliana point to a modified regulation of these genes in the halophyte, possibly through ABA- and/or ethylene-dependent pathways.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Brassicaceae/genética , NADPH Oxidases/genética , Tolerância ao Sal/genética , Arabidopsis/metabolismo , Brassicaceae/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Folhas de Planta/genética , Folhas de Planta/metabolismo , Regiões Promotoras Genéticas/genética , Espécies Reativas de Oxigênio/metabolismo , Salinidade , Plantas Tolerantes a Sal/genética , Estresse Fisiológico/genética
20.
Int J Mol Sci ; 22(19)2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34639024

RESUMO

With no lysine (K) (WNK) kinases comprise a family of serine/threonine kinases belonging to an evolutionary branch of the eukaryotic kinome. These special kinases contain a unique active site and are found in a wide range of eukaryotes. The model plant Arabidopsis has been reported to have 11 WNK members, of which WNK8 functions as a negative regulator of abscisic acid (ABA) signaling. Here, we found that the expression of WNK8 is post-transcriptionally regulated through an upstream open reading frame (uORF) found in its 5' untranslated region (5'-UTR). This uORF has been predicted to encode a conserved peptide named CPuORF58 in both monocotyledons and dicotyledons. The analysis of the published ribosome footprinting studies and the study of the frameshift CPuORF58 peptide with altered repression capability suggested that this uORF causes ribosome stalling. Plants transformed with the native WNK8 promoter driving WNK8 expression were comparable with wild-type plants, whereas the plants transformed with a similar construct with mutated CPuORF58 start codon were less sensitive to ABA. In addition, WNK8 and its downstream target RACK1 were found to synergistically coordinate ABA signaling rather than antagonistically modulating glucose response and flowering in plants. Collectively, these results suggest that the WNK8 expression must be tightly regulated to fulfill the demands of ABA response in plants.


Assuntos
Ácido Abscísico/metabolismo , Proteínas de Arabidopsis/genética , Arabidopsis/genética , Fases de Leitura Aberta , Biossíntese de Proteínas , Proteínas Serina-Treonina Quinases/genética , Regiões 5' não Traduzidas , Sequência de Aminoácidos , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Evolução Molecular , Regulação da Expressão Gênica de Plantas , Germinação/genética , Desenvolvimento Vegetal/genética , Proteínas Serina-Treonina Quinases/metabolismo , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Transdução de Sinais , Sintenia
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