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1.
PLoS Pathog ; 16(8): e1008801, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32866183

RESUMO

Rice stripe virus (RSV) is one of the most destructive viral diseases affecting rice production. However, so far, only one RSV resistance gene has been cloned, the molecular mechanisms underlying host-RSV interaction are still poorly understood. Here, we show that increasing levels or signaling of brassinosteroids (BR) and jasmonic acid (JA) can significantly enhance the resistance against RSV. On the contrary, plants impaired in BR or JA signaling are more susceptible to RSV. Moreover, the enhancement of RSV resistance conferred by BR is impaired in OsMYC2 (a key positive regulator of JA response) knockout plants, suggesting that BR-mediated RSV resistance requires active JA pathway. In addition, we found that RSV infection suppresses the endogenous BR levels to increase the accumulation of OsGSK2, a key negative regulator of BR signaling. OsGSK2 physically interacts with OsMYC2, resulting in the degradation of OsMYC2 by phosphorylation and reduces JA-mediated defense to facilitate virus infection. These findings not only reveal a novel molecular mechanism mediating the crosstalk between BR and JA in response to virus infection and deepen our understanding about the interaction of virus and plants, but also suggest new effective means of breeding RSV resistant crops using genetic engineering.


Assuntos
Brassinosteroides/metabolismo , Ciclopentanos/metabolismo , Oryza , Oxilipinas/metabolismo , Plantas Geneticamente Modificadas , Transdução de Sinais , Tenuivirus , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Oryza/genética , Oryza/metabolismo , Oryza/virologia , Doenças das Plantas/genética , Doenças das Plantas/virologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/virologia , Tenuivirus/genética , Tenuivirus/metabolismo
2.
PLoS Pathog ; 16(8): e1008780, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32866188

RESUMO

Ubiquitin like protein 5 (UBL5) interacts with other proteins to regulate their function but differs from ubiquitin and other UBLs because it does not form covalent conjugates. Ubiquitin and most UBLs mediate the degradation of target proteins through the 26S proteasome but it is not known if UBL5 can also do that. Here we found that the UBL5s of rice and Nicotiana benthamiana interacted with rice stripe virus (RSV) p3 protein. Silencing of NbUBL5s in N. benthamiana facilitated RSV infection, while UBL5 overexpression conferred resistance to RSV in both N. benthamiana and rice. Further analysis showed that NbUBL5.1 impaired the function of p3 as a suppressor of silencing by degrading it through the 26S proteasome. NbUBL5.1 and OsUBL5 interacted with RPN10 and RPN13, the receptors of ubiquitin in the 26S proteasome. Furthermore, silencing of NbRPN10 or NbRPN13 compromised the degradation of p3 mediated by NbUBL5.1. Together, the results suggest that UBL5 mediates the degradation of RSV p3 protein through the 26S proteasome, a previously unreported plant defense strategy against RSV infection.


Assuntos
Proteínas de Plantas/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteólise , Proteínas Repressoras/metabolismo , Tenuivirus/metabolismo , Tabaco/metabolismo , Ubiquitinas/metabolismo , Proteínas Virais/metabolismo , Proteínas de Plantas/genética , Complexo de Endopeptidases do Proteassoma/genética , Proteínas Repressoras/genética , Tenuivirus/genética , Tabaco/genética , Ubiquitinas/genética , Proteínas Virais/genética
3.
Gene ; 762: 145104, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-32889060

RESUMO

Chalcone synthase (CHS, EC 2.3.1.74) is one of the key and rate-limiting enzymes of phenylpropanoid pathway which plays superior roles in the production of secondary metabolites. In the present study a full-length cDNA of CHS gene was isolated and characterized from Coelogyne ovalis, an orchid of ornamental and medicinal importance. The CHS gene sequence from C. ovalis (CoCHS) was found to be 1445 bp and comprised an open reading frame of 1182 bp, encoding for 394 amino acid residues. Further, the sequence alignment and phylogenetic analysis revealed that CoCHS protein shared high degree of similarity with CHS protein of other orchid species. It also confirmed that it contained all four motifs (I to IV) and signature sequence for the functionality of this gene. Structural modeling of CoCHS based on the crystallographic structure of Freesia hybrida indicated that CoCHS had a similar structure. Quantitative polymerase chain reaction (qPCR) disclosed that CoCHS was expressed in all tissues examined, with the highest transcript being in leaves, followed by pseudobulbs and roots. CoCHS expression was also evaluated in the in vitro-raised plantlets under the abiotic stress (dark, cold, UV-B, wounding, salinity). mRNA transcript expression of CHS gene was found to be positively enhanced and regulated by the different stress types. A correlation between the CoCHS transcript expression with flavonoid and anthocyanin contents revealed that a positive correlation existed between metabolites' content and CoCHS expression within the in vivo as well as in the in vitro-raised plant parts.


Assuntos
Aciltransferases/genética , Regulação da Expressão Gênica de Plantas , Orchidaceae/genética , Proteínas de Plantas/genética , Aciltransferases/química , Aciltransferases/metabolismo , Clonagem Molecular , Orchidaceae/classificação , Orchidaceae/metabolismo , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Estresse Fisiológico
4.
PLoS One ; 15(8): e0236226, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32866160

RESUMO

Amine oxidases (AOs) including copper containing amine oxidases (CuAOs) and FAD-dependent polyamine oxidases (PAOs) are associated with polyamine catabolism in the peroxisome, apoplast and cytoplasm and play an essential role in growth and developmental processes and response to biotic and abiotic stresses. Here, we identified PAO genes in common wheat (Triticum aestivum), T. urartu and Aegilops tauschii and reported the genome organization, evolutionary features and expression profiles of the wheat PAO genes (TaPAO). Expression analysis using publicly available RNASeq data showed that TaPAO genes are expressed redundantly in various tissues and developmental stages. A large percentage of TaPAOs respond significantly to abiotic stresses, especially temperature (i.e. heat and cold stress). Some TaPAOs were also involved in response to other stresses such as powdery mildew, stripe rust and Fusarium infection. Overall, TaPAOs may have various functions in stress tolerances responses, and play vital roles in different tissues and developmental stages. Our results provided a reference for further functional investigation of TaPAO proteins.


Assuntos
Resposta ao Choque Frio/genética , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Proteínas de Plantas/genética , Termotolerância/genética , Triticum/genética , Aegilops/enzimologia , Aegilops/genética , Processamento Alternativo , Sequência de Aminoácidos , Conjuntos de Dados como Assunto , Evolução Molecular , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Genoma de Planta , Estudo de Associação Genômica Ampla , Cadeias de Markov , Modelos Genéticos , Peso Molecular , Família Multigênica , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/química , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/metabolismo , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Domínios Proteicos/genética , RNA-Seq , Alinhamento de Sequência , Triticum/enzimologia
5.
Nat Commun ; 11(1): 4393, 2020 09 02.
Artigo em Inglês | MEDLINE | ID: mdl-32879321

RESUMO

Rcr3 is a secreted protease of tomato that is targeted by fungal effector Avr2, a secreted protease inhibitor of the fungal pathogen Cladosporium fulvum. The Avr2-Rcr3 complex is recognized by receptor-like protein Cf-2, triggering hypersensitive cell death (HR) and disease resistance. Avr2 also targets Rcr3 paralog Pip1, which is not required for Avr2 recognition but contributes to basal resistance. Thus, Rcr3 acts as a guarded decoy in this interaction, trapping the fungus into a recognition event. Here we show that Rcr3 evolved > 50 million years ago (Mya), whereas Cf-2 evolved <6Mya by co-opting the pre-existing Rcr3 in the Solanum genus. Ancient Rcr3 homologs present in tomato, potato, eggplants, pepper, petunia and tobacco can be inhibited by Avr2 with the exception of tobacco Rcr3. Four variant residues in Rcr3 promote Avr2 inhibition, but the Rcr3 that co-evolved with Cf-2 lacks three of these residues, indicating that the Rcr3 co-receptor is suboptimal for Avr2 binding. Pepper Rcr3 triggers HR with Cf-2 and Avr2 when engineered for enhanced inhibition by Avr2. Nicotiana benthamiana (Nb) is a natural null mutant carrying Rcr3 and Pip1 alleles with deleterious frame-shift mutations. Resurrected NbRcr3 and NbPip1 alleles were active proteases and further NbRcr3 engineering facilitated Avr2 inhibition, uncoupled from HR signalling. The evolution of a receptor co-opting a conserved pathogen target contrasts with other indirect pathogen recognition mechanisms.


Assuntos
Cladosporium , Resistência à Doença/genética , Peptídeo Hidrolases/genética , Imunidade Vegetal/genética , Solanum , Tabaco , Cladosporium/genética , Cladosporium/metabolismo , Cladosporium/patogenicidade , Evolução Molecular , Proteínas Fúngicas/metabolismo , Genes de Plantas , Interações Hospedeiro-Parasita , Peptídeo Hidrolases/metabolismo , Filogenia , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Inibidores de Proteases/metabolismo , Solanum/genética , Solanum/metabolismo , Solanum/microbiologia , Tabaco/genética , Tabaco/metabolismo , Tabaco/microbiologia
6.
Nat Commun ; 11(1): 4382, 2020 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-32873802

RESUMO

Fusarium graminearum is a causal agent of Fusarium head blight (FHB) and a deoxynivalenol (DON) producer. In this study, OSP24 is identified as an important virulence factor in systematic characterization of the 50 orphan secreted protein (OSP) genes of F. graminearum. Although dispensable for growth and initial penetration, OSP24 is important for infectious growth in wheat rachis tissues. OSP24 is specifically expressed during pathogenesis and its transient expression suppresses BAX- or INF1-induced cell death. Osp24 is translocated into plant cells and two of its 8 cysteine-residues are required for its function. Wheat SNF1-related kinase TaSnRK1α is identified as an Osp24-interacting protein and shows to be important for FHB resistance in TaSnRK1α-overexpressing or silencing transgenic plants. Osp24 accelerates the degradation of TaSnRK1α by facilitating its association with the ubiquitin-26S proteasome. Interestingly, TaSnRK1α also interacts with TaFROG, an orphan wheat protein induced by DON. TaFROG competes against Osp24 for binding with the same region of TaSnRKα and protects it from degradation. Overexpression of TaFROG stabilizes TaSnRK1α and increases FHB resistance. Taken together, Osp24 functions as a cytoplasmic effector by competing against TaFROG for binding with TaSnRK1α, demonstrating the counteracting roles of orphan proteins of both host and fungal pathogens during their interactions.


Assuntos
Proteínas Fúngicas/metabolismo , Fusarium/patogenicidade , Doenças das Plantas/imunologia , Proteínas de Plantas/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Triticum/microbiologia , Fatores de Virulência/metabolismo , Resistência à Doença , Fusarium/imunologia , Fusarium/metabolismo , Interações Hospedeiro-Patógeno/imunologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/imunologia , Plantas Geneticamente Modificadas , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/imunologia , Proteólise , Tricotecenos/metabolismo , Triticum/imunologia
7.
Nat Commun ; 11(1): 4859, 2020 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-32978401

RESUMO

Cell death is intrinsically linked with immunity. Disruption of an immune-activated MAPK cascade, consisting of MEKK1, MKK1/2, and MPK4, triggers cell death and autoimmunity through the nucleotide-binding leucine-rich repeat (NLR) protein SUMM2 and the MAPK kinase kinase MEKK2. In this study, we identify a Catharanthus roseus receptor-like kinase 1-like (CrRLK1L), named LETUM2/MEDOS1 (LET2/MDS1), and the glycosylphosphatidylinositol (GPI)-anchored protein LLG1 as regulators of mekk1-mkk1/2-mpk4 cell death. LET2/MDS1 functions additively with LET1, another CrRLK1L, and acts genetically downstream of MEKK2 in regulating SUMM2 activation. LET2/MDS1 complexes with LET1 and promotes LET1 phosphorylation, revealing an intertwined regulation between different CrRLK1Ls. LLG1 interacts with the ectodomain of LET1/2 and mediates LET1/2 transport to the plasma membrane, corroborating its function as a co-receptor of LET1/2 in the mekk1-mkk1/2-mpk4 cell death pathway. Thus, our data suggest that a trimeric complex consisting of two CrRLK1Ls LET1, LET2/MDS1, and a GPI-anchored protein LLG1 that regulates the activation of NLR SUMM2 for initiating cell death and autoimmunity.


Assuntos
Autoimunidade/genética , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Proteínas Ligadas por GPI/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Autoimunidade/fisiologia , Proteínas de Transporte/imunologia , Catharanthus/genética , Catharanthus/metabolismo , Morte Celular/genética , Proteínas Ligadas por GPI/genética , Regulação da Expressão Gênica de Plantas , Glicosilfosfatidilinositóis , MAP Quinase Quinase Quinases/genética , Sistema de Sinalização das MAP Quinases , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Fosforilação , Proteínas de Plantas/imunologia , Plantas Geneticamente Modificadas , Interferência de RNA , Transcriptoma
8.
Ecotoxicol Environ Saf ; 202: 110904, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32800239

RESUMO

Cation diffusion facilitators (CDFs) play central roles in metal homeostasis and tolerance in plants, but the specific functions of Camellia sinensis CDF-encoding genes and the underlying mechanisms remain unknown. Previously, transcriptome sequencing results in our lab indicated that the expression of CsMTP8.2 in tea plant shoots was down-regulated exposed to excessive amount of Mn2+ conditions. To elucidate the possible mechanisms involved, we systematically identified 13 C. sinensis CsMTP genes from three subfamilies and characterized their phylogeny, structures, and the features of the encoded proteins. The transcription of CsMTP genes was differentially regulated in C. sinensis shoots and roots in responses to high concentrations of Mn, Zn, Fe, and Al. Differences in the cis-acting regulatory elements in the CsMTP8.1 and CsMTP8.2 promoters suggested the expression of these two genes may be differentially regulated. Transient expression analysis indicated that CsMTP8.2 was localized to the plasma membrane in tobacco and onion epidermal cells. Moreover, when heterologously expressed in yeast, CsMTP8.2 conferred tolerance to Ni and Mn but not to Zn. Additionally, heterologous expression of CsMTP8.2 in Arabidopsis thaliana revealed that CsMTP8.2 positively regulated the response to manganese toxicity by decreasing the accumulation of Mn in plants. However, there was no difference in the accumulation of other metals, including Cu, Fe, and Zn. These results suggest that CsMTP8.2 is a Mn-specific transporter that contributes to the efflux of excess Mn2+ from plant cells.


Assuntos
Camellia sinensis/genética , Manganês/toxicidade , Poluentes do Solo/toxicidade , Arabidopsis/metabolismo , Membrana Celular/metabolismo , Regulação da Expressão Gênica de Plantas , Manganês/metabolismo , Filogenia , Células Vegetais/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/metabolismo , Brotos de Planta/metabolismo , Saccharomyces cerevisiae/metabolismo , Chá
9.
Nat Commun ; 11(1): 3797, 2020 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-32732998

RESUMO

Receptor-mediated perception of surface-exposed carbohydrates like lipo- and exo-polysaccharides (EPS) is important for non-self recognition and responses to microbial associated molecular patterns in mammals and plants. In legumes, EPS are monitored and can either block or promote symbiosis with rhizobia depending on their molecular composition. To establish a deeper understanding of receptors involved in EPS recognition, we determined the structure of the Lotus japonicus (Lotus) exopolysaccharide receptor 3 (EPR3) ectodomain. EPR3 forms a compact structure built of three putative carbohydrate-binding modules (M1, M2 and LysM3). M1 and M2 have unique ßαßß and ßαß folds that have not previously been observed in carbohydrate binding proteins, while LysM3 has a canonical ßααß fold. We demonstrate that this configuration is a structural signature for a ubiquitous class of receptors in the plant kingdom. We show that EPR3 is promiscuous, suggesting that plants can monitor complex microbial communities though this class of receptors.


Assuntos
Lipopolissacarídeos/metabolismo , Lotus/microbiologia , Lotus/fisiologia , Mesorhizobium/metabolismo , Proteínas de Plantas/metabolismo , Sequência de Aminoácidos , Mesorhizobium/genética , Fixação de Nitrogênio/fisiologia , Proteínas de Plantas/genética , Dobramento de Proteína , Nódulos Radiculares de Plantas/microbiologia , Nódulos Radiculares de Plantas/fisiologia , Simbiose/fisiologia
10.
Gene ; 761: 145043, 2020 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-32777530

RESUMO

Tonoplast Intrinsic Proteins (TIPs) constitute a significant class of the aquaporins. The TIPs control water trade among cytosolic and vacuolar compartments and can also transport glycerol, ammonia, urea, hydrogen peroxide, metals/metalloids, and so forth. Additionally, TIPs are engaged with different abiotic stress responses and developmental processes like leaf expansion, root elongation and seed germination. In this study, ten TIP genes in the rice genome were identified from Oryza sativa ssp indica. Among these, representative groups of TIP genes were cloned and sequenced whilst some TIP sequences showed stop codons in the coding region. The secondary structure analysis represented six conserved transmembrane helices along with the inter-helical regions having conserved motifs. The representative three-dimensional tetrameric design of protein sequence of TIP1;1 displayed key features like NPA motifs, aromatic/arginine (ar/R) selectivity filters, and Froger's residues. The vacuolar localization, transmembrane topological properties, and conserved motif analysis of the cloned genes altogether supported their identity as TIPs. An unrooted phylogenetic tree delineated the relatedness of TIPs from Oryza with different species and bunched them into five clades. The promoter analysis uncovered key regulons associated with administering abiotic stress responses. Gene expression studies showed thatTIPsare differentially regulated under salt and drought stress at various time points in shoots and roots of rice. Also, the pattern of expression was found to be significantly variable in five different rice tissues. The heat-map based tissue and stress- specific expression analysis supported the experimental findings. In conclusion, the identification and transcript-level expression studies of TIPs significantly contribute towards the comprehension of their utilitarian significance in the abiotic stress response.


Assuntos
Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Oryza/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Aquaporinas/genética , Aquaporinas/metabolismo , Clonagem Molecular/métodos , Secas , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas/genética , Oryza/metabolismo , Filogenia , Folhas de Planta/metabolismo , Cloreto de Sódio/metabolismo , Estresse Fisiológico/genética , Vacúolos/genética , Água/metabolismo
11.
Gene ; 762: 145026, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-32781193

RESUMO

Cannabis has been cultivated for millennia for medicinal, industrial and recreational uses. Our long-term goal is to compare the transcriptomes of cultivars with different cannabinoid profiles for therapeutic purposes. Here we describe the de novo assembly, annotation and initial analysis of two cultivars of Cannabis, a high THC variety and a CBD plus THC variety. Cultivars were grown under different lighting conditions; flower buds were sampled over 71 days. Cannabinoid profiles were determined by ESI-LC/MS. RNA samples were sequenced using the HiSeq4000 platform. Transcriptomes were assembled using the DRAP pipeline and annotated using the BLAST2GO pipeline and other tools. Each transcriptome contained over twenty thousand protein encoding transcripts with ORFs and flanking sequence. Identification of transcripts for cannabinoid pathway and related enzymes showed full-length ORFs that align with the draft genomes of the Purple Kush and Finola cultivars. Two transcripts were found for olivetolic acid cyclase (OAC) that mapped to distinct locations on the Purple Kush genome suggesting multiple genes for OAC are expressed in some cultivars. The ability to make high quality annotated reference transcriptomes in Cannabis or other plants can promote rapid comparative analysis between cultivars and growth conditions in Cannabis and other organisms without annotated genome assemblies.


Assuntos
Canabinoides/biossíntese , Cannabis/genética , Transcriptoma , Cannabis/classificação , Cannabis/metabolismo , Transferases Intramoleculares/genética , Transferases Intramoleculares/metabolismo , Anotação de Sequência Molecular , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
12.
Gene ; 762: 145015, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-32783994

RESUMO

Phosphoenolpyruvate carboxylase (PEPC) is a carbon fixation enzyme which probably plays crucial roles in seed development. A greater number of PEPC isoforms are encoded in the soybean genome, while most of the PEPC isoforms are functionally unknown. In this study, we investigated on soybean PEPC expressed in the external layer of seed coat (ELSC) during seed formation. PEPC activity in ELSC ranged from 0.24 to 1.0 U/g F.W., which could be comparable to those in whole seeds at U per dry matter. Public RNA-Seq data in separated soybean seed tissues revealed that six plant-type PEPC isogenes were substantially expressed in ELSC, and Gmppc1 and Gmppc7 were highly expressed in hourglass cells of ELSC. Gene Ontology enrichment of co-expressed genes with Gmppc1 and Gmppc7 implicated a role of these isogenes in assisting energy production and cellulose biosynthesis. Comparison of PEPC sequences from 16 leguminous species hypothesized adaptive evolution of the Gmppc1 and Gmppc7 lineage after divergence from the other plant-type PEPC lineages. Molecular diversification of these plant-type PEPC was possibly accomplished by adaptation to the functions of the soybean seed tissues. This study indicates that energy demand in immature seeds may be a driving force for the molecular evolution of PEPC.


Assuntos
Fosfoenolpiruvato Carboxilase/genética , Proteínas de Plantas/genética , Soja/genética , Evolução Molecular , Fosfoenolpiruvato Carboxilase/metabolismo , Proteínas de Plantas/metabolismo , Sementes/genética , Sementes/metabolismo , Soja/metabolismo
13.
PLoS One ; 15(8): e0237569, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32817667

RESUMO

Several 'super-complexes' of individual hetero-oligomeric membrane protein complexes, whose function is to facilitate intra-membrane electron and proton transfer and harvesting of light energy, have been previously characterized in the mitochondrial cristae and chloroplast thylakoid membranes. We report the presence of an intra-membrane super-complex dominated by the ATP-synthase, photosystem I (PSI) reaction-center complex and the ferredoxin-NADP+ Reductase (FNR) in the thylakoid membrane. The presence of the super-complex has been documented by mass spectrometry, clear-native PAGE and Western Blot analyses. This is the first documented presence of ATP synthase in a super-complex with the PSI reaction-center located in the non-appressed stromal domain of the thylakoid membrane.


Assuntos
Cloroplastos/metabolismo , Ferredoxina-NADP Redutase/metabolismo , Óxido Nítrico Sintase/metabolismo , Complexo de Proteína do Fotossistema I/metabolismo , Complexo de Proteína do Fotossistema II/metabolismo , ATPases Translocadoras de Prótons/metabolismo , Tilacoides/metabolismo , Trifosfato de Adenosina/metabolismo , Transporte de Elétrons , Fotossíntese , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Spinacia oleracea/crescimento & desenvolvimento , Spinacia oleracea/metabolismo
14.
BMC Bioinformatics ; 21(1): 365, 2020 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-32838742

RESUMO

BACKGROUND: The amount of published full-text articles has increased dramatically. Text mining tools configure an essential approach to building biological networks, updating databases and providing annotation for new pathways. PESCADOR is an online web server based on LAITOR and NLProt text mining tools, which retrieves protein-protein co-occurrences in a tabular-based format, adding a network schema. Here we present an HPC-oriented version of PESCADOR's native text mining tool, renamed to LAITOR4HPC, aiming to access an unlimited abstract amount in a short time to enrich available networks, build new ones and possibly highlight whether fields of research have been exhaustively studied. RESULTS: By taking advantage of parallel computing HPC infrastructure, the full collection of MEDLINE abstracts available until June 2017 was analyzed in a shorter period (6 days) when compared to the original online implementation (with an estimated 2 years to run the same data). Additionally, three case studies were presented to illustrate LAITOR4HPC usage possibilities. The first case study targeted soybean and was used to retrieve an overview of published co-occurrences in a single organism, retrieving 15,788 proteins in 7894 co-occurrences. In the second case study, a target gene family was searched in many organisms, by analyzing 15 species under biotic stress. Most co-occurrences regarded Arabidopsis thaliana and Zea mays. The third case study concerned the construction and enrichment of an available pathway. Choosing A. thaliana for further analysis, the defensin pathway was enriched, showing additional signaling and regulation molecules, and how they respond to each other in the modulation of this complex plant defense response. CONCLUSIONS: LAITOR4HPC can be used for an efficient text mining based construction of biological networks derived from big data sources, such as MEDLINE abstracts. Time consumption and data input limitations will depend on the available resources at the HPC facility. LAITOR4HPC enables enough flexibility for different approaches and data amounts targeted to an organism, a subject, or a specific pathway. Additionally, it can deliver comprehensive results where interactions are classified into four types, according to their reliability.


Assuntos
Software , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Bases de Dados Factuais , Proteínas de Plantas/metabolismo , Mapas de Interação de Proteínas , Zea mays/metabolismo
15.
PLoS Genet ; 16(8): e1008691, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32764743

RESUMO

Primary ciliary dyskinesia (PCD) is characterized by chronic airway disease, reduced fertility, and randomization of the left/right body axis. It is caused by defects of motile cilia and sperm flagella. We screened a cohort of affected individuals that lack an obvious axonemal defect for pathogenic variants using whole exome capture, next generation sequencing, and bioinformatic analysis assuming an autosomal recessive trait. We identified one subject with an apparently homozygous nonsense variant [(c.1762C>T), p.(Arg588*)] in the uncharacterized CFAP57 gene. Interestingly, the variant results in the skipping of exon 11 (58 amino acids), which may be due to disruption of an exonic splicing enhancer. In normal human nasal epithelial cells, CFAP57 localizes throughout the ciliary axoneme. Nasal cells from the PCD patient express a shorter, mutant version of CFAP57 and the protein is not incorporated into the axoneme. The missing 58 amino acids include portions of WD repeats that may be important for loading onto the intraflagellar transport (IFT) complexes for transport or docking onto the axoneme. A reduced beat frequency and an alteration in ciliary waveform was observed. Knockdown of CFAP57 in human tracheobronchial epithelial cells (hTECs) recapitulates these findings. Phylogenetic analysis showed that CFAP57 is highly conserved in organisms that assemble motile cilia. CFAP57 is allelic with the BOP2/IDA8/FAP57 gene identified previously in Chlamydomonas reinhardtii. Two independent, insertional fap57 Chlamydomonas mutant strains show reduced swimming velocity and altered waveforms. Tandem mass tag (TMT) mass spectroscopy shows that FAP57 is missing, and the "g" inner dyneins (DHC7 and DHC3) and the "d" inner dynein (DHC2) are reduced, but the FAP57 paralog FBB7 is increased. Together, our data identify a homozygous variant in CFAP57 that causes PCD that is likely due to a defect in the inner dynein arm assembly process.


Assuntos
Axonema/metabolismo , Transtornos da Motilidade Ciliar/genética , Códon sem Sentido , Dineínas/metabolismo , Proteínas/genética , Células 3T3 , Adulto , Animais , Axonema/fisiologia , Células Cultivadas , Chlamydomonas reinhardtii , Cílios/metabolismo , Cílios/fisiologia , Transtornos da Motilidade Ciliar/patologia , Sequência Conservada , Humanos , Masculino , Camundongos , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas/química , Proteínas/metabolismo , Mucosa Respiratória/metabolismo
16.
PLoS Biol ; 18(8): e3000830, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32810128

RESUMO

Plants are attacked by herbivores, which often specialize on different tissues, and in response, have evolved sophisticated resistance strategies that involve different types of chemical defenses frequently targeted to different tissues. Most known phytohormones have been implicated in regulating these defenses, with jasmonates (JAs) playing a pivotal role in complex regulatory networks of signaling interactions, often generically referred to as "cross talk." The newly identified class of phytohormones, strigolactones (SLs), known to regulate the shoot architecture, remain unstudied with regard to plant-herbivore interactions. We explored the role of SL signaling in resistance to a specialist weevil (Trichobaris mucorea) herbivore of the native tobacco, Nicotiana attenuata, that attacks the root-shoot junction (RSJ), the part of the plant most strongly influenced by alterations in SL signaling (increased branching). As SL signaling shares molecular components, such as the core F-box protein MORE AXILLARY GROWTH 2 (MAX2), with another new class of phytohormones, the karrikins (KARs), which promote seed germination and seedling growth, we generated transformed lines, individually silenced in the expression of NaMAX2, DWARF 14 (NaD14: the receptor for SL) and CAROTENOID CLEAVAGE DIOXYGENASE 7 (NaCCD7: a key enzyme in SL biosynthesis), and KARRIKIN INSENSITIVE 2 (NaKAI2: the KAR receptor). The mature stems of all transgenic lines impaired in the SL, but not the KAR signaling pathway, overaccumulated anthocyanins, as did the stems of plants attacked by the larvae of weevil, which burrow into the RSJs to feed on the pith of N. attenuata stems. T. mucorea larvae grew larger in the plants silenced in the SL pathway, but again, not in the KAI2-silenced plants. These phenotypes were associated with elevated JA and auxin (indole-3-acetic acid [IAA]) levels and significant changes in the accumulation of defensive compounds, including phenolamides and nicotine. The overaccumulation of phenolamides and anthocyanins in the SL pathway-silenced plants likely resulted from antagonism between the SL and JA pathway in N. attenuata. We show that the repressors of SL signaling, suppressor of max2-like (NaSMXL6/7), and JA signaling, jasmonate zim-domain (NaJAZs), physically interact, promoting NaJAZb degradation and releasing JASMONATE INSENSITIVE 1 (JIN1/MYC2) (NaMYC2), a critical transcription factor promoting JA responses. However, the increased performance of T. mucorea larvae resulted from lower pith nicotine levels, which were inhibited by increased IAA levels in SL pathway-silenced plants. This inference was confirmed by decapitation and auxin transport inhibitor treatments that decreased pith IAA and increased nicotine levels. In summary, SL signaling tunes specific sectors of specialized metabolism in stems, such as phenylpropanoid and nicotine biosynthesis, by tailoring the cross talk among phytohormones, including JA and IAA, to mediate herbivore resistance of stems. The metabolic consequences of the interplay of SL, JA, and IAA signaling revealed here could provide a mechanism for the commonly observed pattern of herbivore tolerance/resistance trade-offs.


Assuntos
Herbivoria/fisiologia , Interações Hospedeiro-Parasita , Lactonas/metabolismo , Caules de Planta/metabolismo , Caules de Planta/parasitologia , Transdução de Sinais , Tabaco/metabolismo , Tabaco/parasitologia , Animais , Antocianinas/metabolismo , Ciclopentanos/metabolismo , Ácidos Indolacéticos/metabolismo , Larva , Metabolômica , Oxilipinas/metabolismo , Proteínas de Plantas/metabolismo , Interferência de RNA , Gorgulhos/fisiologia
17.
DNA Cell Biol ; 39(10): 1850-1861, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32790504

RESUMO

Strigolactones (SLs) are the major plant hormones that play important roles in regulating organ development and environmental stress tolerance in plants. Even though the SL-related genes have been identified and well characterized in some plants, the information of SL-related genes in soybean is not fully established yet, especially in response to salt and alkaline stresses. In this study, we identified nine SL biosynthetic genes that include two D27, two CCD7, two CCD8, and three MAX1, as well as seven SL signaling genes that comprised two D14, two MAX2, and three D53 in the soybean genome. We found that SL biosynthetic and signaling genes are evolutionary conserved among different species. Syntenic analysis of these genes revealed their location on nine chromosomes as well as the presence of 10 pairs of duplication genes. Moreover, plant hormone and stress-responsive elements were identified in the promoter regions of SL biosynthetic and signaling genes. By using reverse transcription quantitative real-time PCR, we confirmed that SL genes have different tissue expressions in roots, stems, and leaves. The expression profile of SL biosynthetic and signaling genes under salt and alkaline stresses further confirmed the regulatory roles of SL biosynthetic and signaling genes under stress. In conclusion, we identified and provided valuable information on the soybean SL biosynthetic and signaling genes, and established a foundation for further functional analysis of soybean SL-related genes in response to salt and alkaline stresses.


Assuntos
Compostos Heterocíclicos com 3 Anéis/metabolismo , Lactonas/metabolismo , Proteínas de Plantas/genética , Estresse Salino , Soja/genética , Cromossomos de Plantas/genética , Duplicação Gênica , Proteínas de Plantas/metabolismo , Transdução de Sinais , Soja/metabolismo , Sintenia
18.
PLoS One ; 15(8): e0237845, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32813721

RESUMO

Aluminum (Al3+) toxicity is one of the most important limitations to agricultural production worldwide. The overall response of plants to Al3+ stress has been documented, but the contribution of protein phosphorylation to Al3+ detoxicity and tolerance in plants is unclear. Using a combination of tandem mass tag (TMT) labeling, immobilized metal affinity chromatography (IMAC) enrichment and liquid chromatography-tandem mass spectrometry (LC-MS/MS), Al3+-induced phosphoproteomic changes in roots of Tamba black soybean (TBS) were investigated in this study. The Data collected in this study are available via ProteomeXchange with the identifier PXD019807. After the Al3+ treatment, 189 proteins harboring 278 phosphosites were significantly changed (fold change > 1.2 or < 0.83, p < 0.05), with 88 upregulated, 96 downregulated and 5 up-/downregulated. Enrichment and protein interaction analyses revealed that differentially phosphorylated proteins (DPPs) under the Al3+ treatment were mainly related to G-protein-mediated signaling, transcription and translation, transporters and carbohydrate metabolism. Particularly, DPPs associated with root growth inhibition or citric acid synthesis were identified. The results of this study provide novel insights into the molecular mechanisms of TBS post-translational modifications in response to Al3+ stress.


Assuntos
Alumínio/toxicidade , Fosfoproteínas/metabolismo , Proteínas de Plantas/metabolismo , Proteômica , Soja/metabolismo , Citratos/metabolismo , Fosforilação/efeitos dos fármacos , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismo , Biossíntese de Proteínas/efeitos dos fármacos , Mapas de Interação de Proteínas/efeitos dos fármacos , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Soja/efeitos dos fármacos , Estresse Fisiológico/efeitos dos fármacos , Transcrição Genética/efeitos dos fármacos
19.
PLoS One ; 15(8): e0237884, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32841243

RESUMO

The Solanum tuberosum plant specific insert (StPSI) has a defensive role in potato plants, with the requirements of acidic pH and anionic lipids. The StPSI contains a set of three highly conserved disulfide bonds that bridge the protein's helical domains. Removal of these bonds leads to enhanced membrane interactions. This work examined the effects of their sequential removal, both individually and in combination, using all-atom molecular dynamics to elucidate the role of disulfide linkages in maintaining overall protein tertiary structure. The tertiary structure was found to remain stable at both acidic (active) and neutral (inactive) pH despite the removal of disulfide linkages. The findings include how the dimer structure is stabilized and the impact on secondary structure on a residue-basis as a function of disulfide bond removal. The StPSI possesses an extensive network of inter-monomer hydrophobic interactions and intra-monomer hydrogen bonds, which is likely the key to the stability of the StPSI by stabilizing local secondary structure and the tertiary saposin-fold, leading to a robust association between monomers, regardless of the disulfide bond state. Removal of disulfide bonds did not significantly impact secondary structure, nor lead to quaternary structural changes. Instead, disulfide bond removal induces regions of amino acids with relatively higher or lower variation in secondary structure, relative to when all the disulfide bonds are intact. Although disulfide bonds are not required to preserve overall secondary structure, they may have an important role in maintaining a less plastic structure within plant cells in order to regulate membrane affinity or targeting.


Assuntos
Dissulfetos/metabolismo , Simulação de Dinâmica Molecular , Proteínas de Plantas/metabolismo , Saposinas/metabolismo , Solanum tuberosum/metabolismo , Cisteína/metabolismo , Ligação de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Proteínas de Plantas/química , Multimerização Proteica , Estabilidade Proteica , Estrutura Secundária de Proteína , Sais/química , Enxofre/metabolismo
20.
Ecotoxicol Environ Saf ; 204: 111086, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32781345

RESUMO

In the present research, Silene vulgaris as a representative species growing on both unpolluted and heavy metal (HM) polluted terrains were used to identify ecotype-specific responses to metallic stress. Growth, cell ultrastructure and element accumulations were compared between non-metallicolous (NM), calamine (CAL) and serpentine (SER) specimens untreated with HMs and treated with Pb, Cd and Zn ions under in vitro conditions. Moreover, proteins' modifications related to their level, carbonylation and degradations via vacuolar proteases were verified and linked with potential mechanisms to cope with ions toxicity. Our experiment revealed diversified strategy of HM uptake in NM and both metallicolous ecotypes, in which antagonistic relationship of Zn and Pb/Cd ions provided survival benefits for the whole organism. Despite this similarity, growth rate and metabolic pathways induced in CAL and SER shoots varied significantly. Exposition to HMs in CAL culture led to drop in protein level by approximately 16% compared to the control. This parameter nearly correlated with the enhanced activity of proteases at pH 5.2 as well as possible glutamate changes to proline and reduced glutathione, resulting in intensified growth and first signs of cell senescence. In turn, SER shoots were characterized by growth retardation (to 53% of the control), although protein level and carbonylation were not modified, while a deeper insight into protein network showed its remodeling towards production of polyamines and 2-oxoglutarate delivered to the Krebs cycle. Contrary, an uncontrolled HM influx in NM shoots contributed to morpho-structural disorders accompanied by an increase activity of proteases involved in the degradation of oxidized proteins, what pointed to metal-induced autophagy. Taken together, S. vulgaris ecotypes respond to stress by triggering various mechanisms engaged their survival and/or death under HM treatment.


Assuntos
Cádmio/toxicidade , Chumbo/toxicidade , Proteínas de Plantas/metabolismo , Silene/efeitos dos fármacos , Poluentes do Solo/toxicidade , Zinco/toxicidade , Autofagia/efeitos dos fármacos , Bioacumulação/efeitos dos fármacos , Cádmio/metabolismo , Ecótipo , Glutationa/metabolismo , Chumbo/metabolismo , Modelos Teóricos , Estresse Oxidativo/efeitos dos fármacos , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Brotos de Planta/ultraestrutura , Silene/crescimento & desenvolvimento , Silene/metabolismo , Silene/ultraestrutura , Poluentes do Solo/metabolismo , Zinco/metabolismo
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