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1.
J Fungi (Basel) ; 10(4)2024 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-38667920

RESUMO

Penicillium digitatum is a major postharvest pathogen that threatens the global citrus fruit industry and causes great economic losses annually. In the present study, inhibitory properties of cinnamon bark oil (CBO) against P. digitatum in vitro were investigated. Results indicated that 0.03% CBO could efficiently inhibit the spore germination, germ tube elongation, mycelial growth, colonial expansion and conidial accumulation of P. digitatum. The results of fluorescein diacetate (FDA) and MitoTraker Orange (MTO) staining also proved the suppression effects of CBO against P. digitatum. Meanwhile, CBO could inhibit green mold rots induced by P. digitatum in citrus fruit when the working concentration of CBO exceeded 0.06%. In addition, the expressions of 12 genes critical for the growth and virulence of P. digitatum were also significantly regulated under CBO stress. Through a transcriptomic analysis, a total of 1802 common differentially expressed genes (DEGs) were detected in P. digitatum after 4 h and 8 h of CBO treatment. Most of the DEG products were associated with carbohydrate, amino acid and lipid metabolism. They directly or indirectly led to the disturbance of the membrane and the generation of reactive oxygen species (ROS). Our results may deepen the understanding of antifungal properties of CBO against P. digitatum and provide the theoretical foundation to uncover the antifungal mechanism of CBO at the molecular level.

2.
J Fungi (Basel) ; 9(10)2023 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-37888255

RESUMO

Penicillium expansum is the most popular post-harvest pathogen and causes blue mold disease in pome fruit and leads to significant economic losses worldwide every year. However, the fundamental regulation mechanisms of growth in P. expansum are unclear. Recently, non-coding RNAs (ncRNAs) have attracted more attention due to critical roles in normalizing gene expression and maintaining cellular genotypes in organisms. However, the research related to ncRNAs in P. expansum have not been reported. Therefore, to provide an overview of ncRNAs on composition, distribution, expression changes, and potential targets in the growth process, a comparative transcriptomic analysis was performed on spores and mycelia of P. expansum in the present study. A total of 2595 novel mRNAs, 3362 long non-coding RNAs (lncRNAs), 10 novel microRNAs (miRNAs), 86 novel small interfering RNAs (siRNAs), and 11,238 circular RNAs (circRNAs) were predicted and quantified. Of these, 1482 novel mRNAs, 5987 known mRNAs, 2047 lncRNAs, 40 miRNAs, 38 novel siRNAs, and 9235 circRNAs were differentially expressed (DE) in response to the different development stages. Afterward, the involved functions and pathways of DE RNAs were revealed via Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) database enrichment analysis. The interaction networks between mRNAs, lncRNAs, and miRNAs were also predicted based on their correlation coefficient of expression profiles. Among them, it was found that miR168 family members may play important roles in fungal growth due to their central location in the network. These findings will contribute to a better understanding on regulation machinery at the RNA level on fungal growth and provide a theoretical basis to develop novel control strategies against P. expansum.

3.
Plants (Basel) ; 11(24)2022 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-36559681

RESUMO

SlSPL-CNR is a multifunctional transcription factor gene that plays important roles in regulating tomato fruit ripening. However, the molecular basis of SlSPL-CNR in the regulatory networks is not exactly clear. In the present study, the biochemical characteristics and expression levels of genes involved in ethylene biosynthesis in Colorless nonripening (Cnr) natural mutant were determined. The proteomic changes during the ripening stage were also uncovered by isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomic analysis. Results indicated that both the lycopene content and soluble solid content (SSC) in Cnr fruit were lower than those in wild-type AC fruit. Meanwhile, pH, flavonoid content, and chlorophyll content were higher in Cnr fruit. Expressions of genes involved in ethylene biosynthesis were also downregulated or delayed in Cnr fruit. Furthermore, 1024 and 1234 differentially expressed proteins (DEPs) were respectively identified for the breaker and 10 days postbreaker stages. Among them, a total of 512 proteins were differentially expressed at both stages. In addition, the functions of DEPs were classified by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Results would lay the groundwork for wider explorations of the regulatory mechanism of SlSPL-CNR on tomato fruit ripening.

4.
J Fungi (Basel) ; 7(2)2021 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-33572180

RESUMO

Penicillium expansum is a major postharvest pathogen that mainly threatens the global pome fruit industry and causes great economic losses annually. In the present study, the antifungal effects and potential mechanism of cinnamon oil against P. expansum were investigated. Results indicated that 0.25 mg L-1 cinnamon oil could efficiently inhibit the spore germination, conidial production, mycelial accumulation, and expansion of P. expansum. In addition, it could effectively control blue mold rots induced by P. expansum in apples. Cinnamon oil could also reduce the expression of genes involved in patulin biosynthesis. Through a proteomic quantitative analysis, a total of 146 differentially expressed proteins (DEPs) involved in the carbohydrate metabolic process, most of which were down-regulated, were noticed for their large number and functional significance. Meanwhile, the expressions of 14 candidate genes corresponding to DEPs and the activities of six key regulatory enzymes (involving in cellulose hydrolyzation, Krebs circle, glycolysis, and pentose phosphate pathway) showed a similar trend in protein levels. In addition, extracellular carbohydrate consumption, intracellular carbohydrate accumulation, and ATP production of P. expansum under cinnamon oil stress were significantly decreased. Basing on the correlated and mutually authenticated results, we speculated that disturbing the fungal carbohydrate metabolic process would be partly responsible for the inhibitory effects of cinnamon oil on P. expansum growth. The findings would provide new insights into the antimicrobial mode of cinnamon oil.

5.
J Exp Bot ; 71(10): 2995-3011, 2020 05 30.
Artigo em Inglês | MEDLINE | ID: mdl-32016417

RESUMO

SlSPL-CNR, an SBP-box transcription factor (TF) gene residing at the epimutant Colourless non-ripening (Cnr) locus, is involved in tomato ripening. This epimutant provides a unique model to investigate the (epi)genetic basis of fruit ripening. Here we report that SlSPL-CNR is a nucleus-localized protein with a distinct monopartite nuclear localization signal (NLS). It consists of four consecutive residues ' 30KRKR33' at the N-terminus of the protein. Mutation of the NLS abolishes SlSPL-CNR's ability to localize in the nucleus. SlSPL-CNR comprises two zinc-finger motifs (ZFMs) within the C-terminal SBP-box domain. Both ZFMs contribute to zinc-binding activity. SlSPL-CNR can induce cell death in tomato and tobacco, dependent on its nuclear localization. However, the two ZFMs have differential impacts on SlSPL-CNR's induction of severe necrosis or mild necrotic ringspot. NLS and ZFM mutants cannot complement Cnr fruits to ripen. SlSPL-CNR interacts with SlSnRK1. Virus-induced SlSnRK1 silencing leads to reduction in expression of ripening-related genes and inhibits ripening in tomato. We conclude that SlSPL-CNR is a multifunctional protein that consists of a distinct monopartite NLS, binds to zinc, and interacts with SlSnRK1 to affect cell death and tomato fruit ripening.


Assuntos
Solanum lycopersicum , Morte Celular , Etilenos , Frutas/genética , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
6.
Plant Sci ; 278: 113-117, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30471724

RESUMO

Non-cell autonomous RNA silencing can spread from cell to cell and over long-distances in animals and plants. This process is genetically determined and requires mobile RNA signals. Genetic requirement and molecular nature of the mobile signals for non-cell-autonomous RNA silencing were intensively investigated in past few decades. No consensus dogma for mobile silencing can be reached in plants, yet published data are sometimes inconsistent and controversial. Thus, the genetic requirements and molecular signals involved in plant mobile silencing are still poorly understood. This article revisits our present understanding of intercellular and systemic non-cell autonomous RNA silencing, and summarises current debates on RNA signals for mobile silencing. In particular, we discuss new evidence on siRNA mobility, a DCL2-dependent genetic network for mobile silencing and its potential biological relevance as well as 22 nt siRNA being a mobile signal for non-cell-autonomous silencing in both Arabidopsis and Nicotiana benthamiana. This sets up a new trend in unravelling genetic components and small RNA signal molecules for mobile silencing in (across) plants and other organisms of different kingdoms. Finally we raise several outstanding questions that need to be addressed in future plant silencing research.


Assuntos
Modelos Genéticos , Plantas/genética , Interferência de RNA , Comunicação Celular
7.
Molecules ; 23(6)2018 06 08.
Artigo em Inglês | MEDLINE | ID: mdl-29890672

RESUMO

Mango peel, the main by-product of juice processing, possesses appreciable quantities of bioactive phenolic compounds and is worthy of further utilization. The present work reports for the first time the HPLC analysis and in vitro antioxidant evaluation of mango peel phenols (MPPs) and their cytotoxic effect on the A549 lung cancer cell line. These results indicated that mango peel has the total phenolic content of 723.2 ± 0.93 mg·kg−1 dry mango peel (DMP), which consisted mainly of vanillic aldehyde, caffeic acid, chlorogenic acid, gallic acid, procyanidin B2 and oleanolic acid. Antioxidant assays showed that MPPs had strong antioxidant activities, with 92 ± 4.2% of DPPH radical scavenging rate, 79 ± 2.5% of ABTS radical inhibition rate and 4.7 ± 0.5 µM Trolox equivalents per kg−1 DMP of ferric reducing power. Gallic acid possess a stronger antioxidant capacity than other phenols. In vitro cytotoxic tests suggested that mango peel extract (MPE) had an IC50 value of 15 mg·mL−1 and MPPs had a stronger inhibitory effect on the A549 cell line. Oleanolic acid exhibited the strongest cytotoxicity, with an IC50 value of 4.7 µM, which was similar with that of the positive control 5-fluorouracil.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Antioxidantes/farmacologia , Mangifera/química , Ácido Oleanólico/farmacologia , Fenóis/farmacologia , Estruturas Vegetais/química , Células A549 , Cromatografia Líquida de Alta Pressão , Humanos , Concentração Inibidora 50
8.
Microbiologyopen ; 7(3): e00562, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29205951

RESUMO

In this study, Penicillium expansum, a common destructive phytopathogen and patulin producer was isolated from naturally infected apple fruits and identified by morphological observation and rDNA-internal transcribed spacer analysis. Subsequently, a global view of the transcriptome and proteome alteration of P. expansum spores during germination was evaluated by RNA-seq (RNA sequencing) and iTRAQ (isobaric tags for relative and absolute quantitation) approaches. A total of 3,026 differentially expressed genes (DEGs), 77 differentially expressed predicted transcription factors and 489 differentially expressed proteins (DEPs) were identified. The next step involved screening out 130 overlapped candidates through correlation analysis between the RNA-seq and iTRAQ datasets. Part of them showed a different expression trend in the mRNA and protein levels, and most of them were involved in metabolism and genetic information processing. These results not only highlighted a set of genes and proteins that were important in deciphering the molecular processes of P. expansum germination but also laid the foundation to develop effective control methods and adequate environmental conditions.


Assuntos
Perfilação da Expressão Gênica , Penicillium/crescimento & desenvolvimento , Penicillium/genética , Proteoma/análise , Esporos Bacterianos/crescimento & desenvolvimento , Esporos Bacterianos/genética , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Frutas/microbiologia , Malus/microbiologia , Microscopia , Penicillium/citologia , Penicillium/isolamento & purificação , Análise de Sequência de DNA
9.
Int J Food Microbiol ; 244: 1-10, 2017 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-28042969

RESUMO

In this study, the effects of exogenous potassium phosphite (Phi) on growth and patulin production of postharvest pathogen Penicillium expansum were assessed. The results indicated that P. expansum under 5mmol/L Phi stress presented obvious development retardation, yield reduction of patulin and lower infectivity to apple fruit. Meanwhile, expression analysis of 15 genes related to patulin biosynthesis suggested that Phi mainly affected the early steps of patulin synthetic route at transcriptional level. Furthermore, a global view of proteome and transcriptome alteration of P. expansum spores during 6h of Phi stress was evaluated by iTRAQ (isobaric tags for relative and absolute quantitation) and RNA-seq (RNA sequencing) approaches. A total of 582 differentially expressed proteins (DEPs) and 177 differentially expressed genes (DEGs) were acquired, most of which participated in carbohydrate metabolism, amino acid metabolism, lipid metabolism, genetic information processing and biosynthesis of secondary metabolites. Finally, 39 overlapped candidates were screened out through correlational analysis between iTRAQ and RNA-seq datasets. These findings will afford more precise and directional clues to explore the inhibitory mechanism of Phi on growth and patulin biosynthesis of P. expansum, and be beneficial to develop effective controlling approaches based on Phi.


Assuntos
Desinfecção/métodos , Fungicidas Industriais/farmacologia , Patulina/biossíntese , Penicillium/crescimento & desenvolvimento , Penicillium/metabolismo , Fosfitos/farmacologia , Compostos de Potássio/farmacologia , Sequência de Bases , Manipulação de Alimentos , Microbiologia de Alimentos , Frutas/microbiologia , Malus/microbiologia , Penicillium/genética , Proteoma/análise , Análise de Sequência de RNA
10.
Int J Food Microbiol ; 241: 1-6, 2017 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-27728853

RESUMO

Anthracnose caused by Colletotrichum gloeosporioides is one of the most important postharvest diseases in mango fruit, often causing huge economic losses. In this study, the effect of 1-methylcyclopropene (1-MCP) against anthracnose in postharvest mango fruit and the mechanisms involved were investigated. 1-MCP induced reactive oxygen species (ROS) generation, damaged the mitochondria and destroyed the integrity of plasma membrane of spores of C. gloeosporioides, significantly suppressing spore germination and mycelial growth of C. gloeosporioides. 1-MCP also decreased the decay incidence and lesion expansion of mango fruit caused by C. gloeosporioides. For the first time this study demonstrated that 1-MCP suppressed anthracnose of postharvest mango fruit by directly inhibiting spore germination and mycelial growth of C. gloeosporioides, thus providing a promising strategy for disease control.


Assuntos
Antifúngicos/química , Membrana Celular/efeitos dos fármacos , Colletotrichum/efeitos dos fármacos , Ciclopropanos/química , Mangifera/microbiologia , Mitocôndrias/efeitos dos fármacos , Antifúngicos/farmacologia , Microbiologia de Alimentos , Frutas/microbiologia , Doenças das Plantas/microbiologia , Espécies Reativas de Oxigênio/metabolismo
11.
Sci Rep ; 5: 9192, 2015 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-25778911

RESUMO

Naturally-occurring epimutants are rare and have mainly been described in plants. However how these mutants maintain their epigenetic marks and how they are inherited remain unknown. Here we report that CHROMOMETHYLASE3 (SlCMT3) and other methyltransferases are required for maintenance of a spontaneous epimutation and its cognate Colourless non-ripening (Cnr) phenotype in tomato. We screened a series of DNA methylation-related genes that could rescue the hypermethylated Cnr mutant. Silencing of the developmentally-regulated SlCMT3 gene results in increased expression of LeSPL-CNR, the gene encodes the SBP-box transcription factor residing at the Cnr locus and triggers Cnr fruits to ripen normally. Expression of other key ripening-genes was also up-regulated. Targeted and whole-genome bisulfite sequencing showed that the induced ripening of Cnr fruits is associated with reduction of methylation at CHG sites in a 286-bp region of the LeSPL-CNR promoter, and a decrease of DNA methylation in differentially-methylated regions associated with the LeMADS-RIN binding sites. Our results indicate that there is likely a concerted effect of different methyltransferases at the Cnr locus and the plant-specific SlCMT3 is essential for sustaining Cnr epi-allele. Maintenance of DNA methylation dynamics is critical for the somatic stability of Cnr epimutation and for the inheritance of tomato non-ripening phenotype.


Assuntos
DNA (Citosina-5-)-Metiltransferases/metabolismo , Epigênese Genética , Mutação , Fenótipo , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Alelos , Metilação de DNA , Etilenos/biossíntese , Frutas , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Inativação Gênica , Estudo de Associação Genômica Ampla , Regiões Promotoras Genéticas , Transdução de Sinais , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
12.
Sci Rep ; 5: 7852, 2015 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-25597857

RESUMO

In plants, microRNAs (miRNAs) play essential roles in growth, development, yield, stress response and interactions with pathogens. However no miRNA has been experimentally documented to be functionally involved in fruit ripening although many miRNAs have been profiled in fruits. Here we show that SlymiR157 and SlymiR156 differentially modulate ripening and softening in tomato (Solanum lycopersicum). SlymiR157 is expressed and developmentally regulated in normal tomato fruits and in those of the Colourless non-ripening (Cnr) epimutant. It regulates expression of the key ripening gene LeSPL-CNR in a likely dose-dependent manner through miRNA-induced mRNA degradation and translation repression. Viral delivery of either pre-SlymiR157 or mature SlymiR157 results in delayed ripening. Furthermore, qRT-PCR profiling of key ripening regulatory genes indicates that the SlymiR157-target LeSPL-CNR may affect expression of LeMADS-RIN, LeHB1, SlAP2a and SlTAGL1. However SlymiR156 does not affect the onset of ripening, but it impacts fruit softening after the red ripe stage. Our findings reveal that working together with a ripening network of transcription factors, SlymiR157 and SlymiR156 form a critical additional layer of regulatory control over the fruit ripening process in tomato.


Assuntos
Frutas/genética , MicroRNAs/biossíntese , Proteínas de Plantas/biossíntese , RNA de Plantas/genética , Solanum lycopersicum/genética , Frutas/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Solanum lycopersicum/crescimento & desenvolvimento , MicroRNAs/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas
13.
J Proteomics ; 103: 47-56, 2014 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-24675182

RESUMO

Penicillium expansum is an important fungal pathogen, which causes blue mold rot in various fruits and produces a mycotoxin (patulin) with potential damage to public health. Here, we found that nitric oxide (NO) donor could significantly inhibit germinability of P. expansum spores, resulting in lower virulence to apple fruit. Based on two dimension electrophoresis (2-DE) and mass spectrometry (MS) analysis, we identified ten differentially expressed proteins in response to exogenous NO in P. expansum. Among of them, five proteins, such as glutamine synthetase (GS), amidohydrolase, nitrilases, nitric oxide dioxygenase (NOD) and heat shock protein 70, were up-regulated. Others including tetratricopeptide repeat domain, UDP-N-acetylglucosamine pyrophosphorylase, enolase (Eno), heat shock protein 60 and K homology RNA-binding domain were down-regulated. The expression of three genes associated with the identified proteins (GS, NOD, and Eno) was evaluated at the mRNA level by RT-PCR. Our results provide the novel evidence for understanding the mechanism, by which NO regulates growth of P. expansum and its virulence. BIOLOGICAL SIGNIFICANCE: Crop diseases caused by fungal pathogens lead to huge economic losses every year in the world. Application of chemical fungicides to control diseases brings the concern about food and environmental safety. Screening new antimicrobial compounds and exploring involved mechanisms have great significance to development of new disease management strategies. Nitric oxide (NO), as an important intracellular signaling molecule, has been proved to be involved in many physiological processes and defense responses during plant-pathogen interactions. In this study, we firstly found that NO at high concentration could distinctly delay spore germination and significantly reduce virulence of P. expansum to fruit host, identified some important proteins in response to NO stress and characterized the functions of these proteins. These results provide novel evidence for understanding the mechanism of NO regulating virulence of the fungal pathogen, but are beneficial for screening new targets of antifungal compounds.


Assuntos
Óxido Nítrico/farmacologia , Penicillium/metabolismo , Esporos Fúngicos/efeitos dos fármacos , Fungicidas Industriais/farmacologia , Malus/microbiologia , Doadores de Óxido Nítrico/farmacologia , Nitroprussiato/farmacologia , Penicillium/efeitos dos fármacos , Doenças das Plantas/prevenção & controle , Proteômica , Esporos Fúngicos/fisiologia , Virulência/efeitos dos fármacos
14.
Plant Signal Behav ; 8(11): e27142, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24305652

RESUMO

Virus-induced gene complementation (VIGC), a plant virus technology based on Potato virus X for transient overexpression of endogenous genes complemented tomato mutants, resulting in non-ripening fruits to ripen. This efficient "gain-of-function" approach involves no stable transformation, and reveals a fruit-specific transcriptional network that may exist among key transcription factors in modulating tomato ripening. Thus, VIGC represents a novel and feasible strategy for gene functional analysis in plants.


Assuntos
Genes de Plantas , Técnicas Genéticas , Potexvirus/fisiologia , Solanum lycopersicum/genética , Solanum lycopersicum/virologia , Solanum lycopersicum/crescimento & desenvolvimento , Mutação/genética , Proteínas de Plantas/metabolismo
15.
PLoS One ; 8(8): e70959, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23990918

RESUMO

In non-climacteric fruits, the respiratory increase is absent and no phytohormone is appearing to be critical for their ripening process. They must remain on the parent plant to enable full ripening and be picked at or near the fully ripe stage to obtain the best eating quality. However, huge losses often occur for their quick post-harvest senescence. To understanding the complex mechanism of non-climacteric fruits post-harvest senescence, we constructed two small RNA libraries and one degradome from strawberry fruit stored at 20°C for 0 and 24 h. A total of 88 known and 1224 new candidate miRNAs, and 103 targets cleaved by 19 known miRNAs families and 55 new candidatemiRNAs were obtained. These targets were associated with development, metabolism, defense response, signaling transduction and transcriptional regulation. Among them, 14 targets, including NAC transcription factor, Auxin response factors (ARF) and Myb transcription factors, cleaved by 6 known miRNA families and 6 predicted candidates, were found to be involved in regulating fruit senescence. The present study provided valuable information for understanding the quick senescence of strawberry fruit, and offered a foundation for studying the miRNA-mediated senescence of non-climacteric fruits.


Assuntos
Fragaria/genética , Frutas/fisiologia , Sequenciamento de Nucleotídeos em Larga Escala/métodos , MicroRNAs/metabolismo , Fragaria/fisiologia , Frutas/genética , Regulação da Expressão Gênica de Plantas , Biblioteca Gênica , Genoma de Planta , RNA de Plantas/genética , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais , Temperatura , Fatores de Tempo , Fatores de Transcrição/metabolismo , Transcrição Gênica
16.
J Proteomics ; 82: 113-29, 2013 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-23474080

RESUMO

In this study, a comparative proteomic analysis was employed to identify fuzz fiber initiation-related proteins in wild-type diploid cotton (Gossypium arboreum L.) and its fuzzless mutant. Temporal changes in global proteomes were examined using 2-DE at five developmental time points for fuzz fiber initiation, and 71 differentially expressed protein species were identified by MS, 45 of which were preferentially accumulated in the wild-type. These proteins were assigned to several functional categories, mainly in cell response/signal transduction, redox homeostasis, protein metabolism and energy/carbohydrate metabolism. It was remarkable that more than ten key proteins with high-abundance were involved in gibberellic acid (GA) signaling and ROS scavenging, and increasing concentrations of active GAs and H2O2 were also detected approximately 5dpa in wild type ovules. Furthermore, in vivo GA and H2O2 treatments of ovules inside young bolls showed that these compounds can synergistically promote fuzz fiber initiation. Our findings not only described a dynamic protein network supporting fuzz initiation in diploid cotton fiber ovules, but also deepened our understanding of the molecular basis of cotton fiber initiation. BIOLOGICAL SIGNIFICANCE: Our study reported the identification of differentially expressed proteins in wild-type diploid cotton (G. arboreum L.) and its fuzzless mutant by comparative proteomic approach. In total, 71 protein species related to fuzz initiation were identified by MS. These proteins were assigned to several functional categories, mainly in energy/carbohydrate metabolism, protein metabolism, signal transduction, redox homeostasis etc. Importantly, a number of key proteins were found to be associated with GA signaling and ROS scavenging. In consistence with these findings, we detected the increase of GAs and H2O2 concentrations during fiber initiation, and our in vivo ovule experiments with GA and H2O2 injection and following microscopy observation of fuzz fiber initiation supported promoting effects of GA and H2O2 on cotton fiber initiation. These findings depicted a dynamic protein network supporting cotton fiber initiation in diploid cotton ovules. Our study is of major significance for understanding the molecular mechanisms controlling fuzz initiation and also provides a solid basis for further functional research of single nodes of this network in relation to cotton fiber initiation.


Assuntos
Regulação da Expressão Gênica de Plantas/fisiologia , Gossypium/metabolismo , Mutação , Proteínas de Plantas/biossíntese , Metabolismo dos Carboidratos/efeitos dos fármacos , Metabolismo dos Carboidratos/fisiologia , Fibra de Algodão , Diploide , Eletroforese em Gel Bidimensional , Metabolismo Energético/efeitos dos fármacos , Metabolismo Energético/fisiologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Giberelinas/farmacologia , Gossypium/genética , Peróxido de Hidrogênio/farmacologia , Oxidantes/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/genética , Proteômica/métodos , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia
17.
Sci Rep ; 2: 836, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23150786

RESUMO

Plant virus technology, in particular virus-induced gene silencing, is a widely used reverse- and forward-genetics tool in plant functional genomics. However the potential of virus technology to express genes to induce phenotypes or to complement mutants in order to understand the function of plant genes is not well documented. Here we exploit Potato virus X as a tool for virus-induced gene complementation (VIGC). Using VIGC in tomato, we demonstrated that ectopic viral expression of LeMADS-RIN, which encodes a MADS-box transcription factor (TF), resulted in functional complementation of the non-ripening rin mutant phenotype and caused fruits to ripen. Comparative gene expression analysis indicated that LeMADS-RIN up-regulated expression of the SBP-box (SQUAMOSA promoter binding protein-like) gene LeSPL-CNR, but down-regulated the expression of LeHB-1, an HD-Zip homeobox TF gene. Our data support the hypothesis that a transcriptional network may exist among key TFs in the modulation of fruit ripening in tomato.


Assuntos
Solanum lycopersicum/metabolismo , Fatores de Transcrição/metabolismo , Frutas/metabolismo , Redes Reguladoras de Genes , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Metionina/metabolismo , Mutação , Fenótipo , Potexvirus/genética , Fatores de Transcrição/genética , Proteínas Virais/genética , Proteínas Virais/metabolismo
18.
Curr Microbiol ; 62(1): 229-34, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20593183

RESUMO

The effects of nitric oxide (NO) on spore germination of Penicillium expansum were investigated and a possible mechanism was evaluated. The results indicated that NO released by sodium nitroprusside (SNP) significantly suppressed fungal growth. With the use of an oxidant sensitive probe and Western blot analysis, an increased level of intracellular reactive oxygen species (ROS) and enhanced carbonylation damage were detected in spores of P. expansum under NO stress. Exogenous superoxide dismutase (SOD) and ascorbic acid (Vc) could increase the resistance of the spore to the inhibitory effect of NO. The activities of SOD and catalase (CAT), as well as ATP content in spores under NO stress were also lower than those in the control. We suggest that NO in high concentration induces the generation of ROS which subsequently causes severe oxidative damage to proteins crucial to the process of spore germination of P. expansum.


Assuntos
Antifúngicos/metabolismo , Óxido Nítrico/metabolismo , Oxidantes/metabolismo , Penicillium/efeitos dos fármacos , Penicillium/crescimento & desenvolvimento , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/crescimento & desenvolvimento , Citoplasma/química , Espécies Reativas de Oxigênio/análise
19.
J Proteome Res ; 9(1): 298-307, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19951004

RESUMO

Spore germination is the first step for fungal pathogens to infect host plants. The pH value, as one of the most important environmental parameters, has critical influence on spore germination. In this study, effects of ambient pH on spore germination were determined by culturing spores of Penicillium expansum in medium with pH values at 2.0, 5.0 and 8.0, and involved mechanisms were further investigated through methods of comparative proteomics. The results demonstrated that spore germination of P. expansum was obviously inhibited at pH 2.0 and 8.0. Using quadrupole time-of-flight tandem mass spectrometer, 34 proteins with significant changes in abundance were identified. Among them, 17 proteins were related to protein synthesis and folding, and most of them were down-regulated at pH 2.0 and 8.0. Accordingly, lower content of total soluble proteins and higher ratio of aggregated proteins were observed in spores at pH 2.0 and 8.0. In addition, it was found that ambient pH could affect intracellular pH and ATP level of P. expansum spores. These findings indicated that ambient pH might affect spore germination of P. expansum by changing intracellular pH and regulating protein expression. Further, impairing synthesis and folding of proteins might be one of the main reasons.


Assuntos
Proteínas Fúngicas/metabolismo , Penicillium/fisiologia , Proteômica/métodos , Estresse Fisiológico/fisiologia , Trifosfato de Adenosina/metabolismo , Meios de Cultura/química , Eletroforese em Gel Bidimensional , Proteínas Fúngicas/biossíntese , Concentração de Íons de Hidrogênio , Penicillium/citologia , Penicillium/metabolismo , Dobramento de Proteína , Multimerização Proteica , Proteoma/metabolismo , Solubilidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Esporos Fúngicos/citologia , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/metabolismo
20.
Plant Cell Physiol ; 50(2): 230-42, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19068492

RESUMO

In this study, we found that oxalic acid (OA) at the concentration of 5 mM could delay jujube fruit sene-scence by reducing ethylene production, repressing fruit reddening and reducing alcohol content, which consequently increased fruit resistance against blue mold caused by Penicillium expansum. In order to gain a further understanding of the mechanism by which OA delays senescence and increases disease resistance of jujube fruit, we used a proteomics approach to compare soluble proteome of jujube fruits treated with water or 5 mM OA for 10 min. A total of 25 differentially expressed proteins were identified by using electrospray ionization quadrupole time-of-flight tandem mass spectrometry (ESI-Q-TOF-MS/MS). Among these proteins, alcohol dehydrogenase 1, which plays a direct role in ethanol metabolism, was repressed, and the abundances of three photosynthesis-related proteins was enhanced in jujube fruit after OA treatment. The protein identified as a cystathionine beta-synthase domain-containing protein, which can regulate ethylene precursors, was also induced by OA treatment. The activity of 1-aminocyclopropane-1-carboxylic acid synthase was significantly suppressed in OA-treated jujube fruit. In addition, three proteins related to the defense/stress response were up-regulated by OA, and contributed to the establishment of systemic resistance induced by OA in jujube fruits. These results indicated that OA treatment might affect ethanol and ethylene metabolism, resulting in delaying senescence, and increase resistance of jujube fruits against fungal pathogens.


Assuntos
Frutas/efeitos dos fármacos , Ácido Oxálico/farmacologia , Proteômica , Ziziphus/efeitos dos fármacos , Álcool Desidrogenase/metabolismo , Clorofila/análise , Etanol/análise , Etilenos/biossíntese , Frutas/metabolismo , Frutas/microbiologia , Liases/metabolismo , Penicillium/patogenicidade , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Ziziphus/metabolismo , Ziziphus/microbiologia
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