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1.
Int J Food Microbiol ; 330: 108713, 2020 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-32512363

RESUMO

Sodium metabisulfite (SMBS), potassium metabisulfite (PMBS), aluminum sulfate (AlS) and aluminum potassium sulfate (AlPS), common sulfur-containing salts used as food additives, were evaluated for their antifungal activity against Penicillium digitatum, Penicillium italicum and Geotrichum citri-aurantii, the most economically important pathogens causing postharvest diseases of citrus fruits. In vitro radial mycelial growth was measured on potato dextrose agar (PDA) Petri dishes amended with five different concentrations of the salts (10, 20, 30, 50, 100 mM) after 7 d of incubation at 25 °C. SMBS and PMBS at all concentrations, and AIS and AIPS above 20 mM, completely inhibited the growth of these fungi. The curative antifungal activity of the four salts to control citrus green (GM) and blue (BM) molds and sour rot (SR) was evaluated on 'Valencia' oranges artificially inoculated in rind wounds with P. digitatum, P. italicum and G. citri-aurantii, respectively. In vivo primary screenings showed no significant antifungal activity of AlS and AlPS to control the three diseases at any dose tested, but SMBS and PMBS reduced the incidence and severity of GM, BM and SR at various concentrations. Effective salts and concentrations were selected for in vivo dip treatments in small-scale trials. Dips at room temperature (20 °C) in SMBS and PMBS at 20 and 50 mM for 60 or 120 s significantly reduced the incidence and severity of GM and BM, with PMBS at 50 mM for 120 s the most effective treatment. Conversely, dips in SMBS and PMBS at 50 mM for 60 or 120 s did not reduce SR incidence and severity. SMBS and PMBS treatments are potentially new tools to be included in reduced-risk non-polluting strategies to control Penicillium diseases, but not SR, on citrus fruits.


Assuntos
Antifúngicos/farmacologia , Citrus/microbiologia , Aditivos Alimentares/farmacologia , Doenças das Plantas/prevenção & controle , Enxofre/farmacologia , Aditivos Alimentares/química , Conservação de Alimentos/métodos , Frutas/microbiologia , Fungos/efeitos dos fármacos , Fungos/crescimento & desenvolvimento , Doenças das Plantas/microbiologia , Temperatura , Fatores de Tempo
2.
PLoS One ; 15(5): e0227396, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32469865

RESUMO

Elsinoë fawcettii, a necrotrophic fungal pathogen, causes citrus scab on numerous citrus varieties around the world. Known pathotypes of E. fawcettii are based on host range; additionally, cryptic pathotypes have been reported and more novel pathotypes are thought to exist. E. fawcettii produces elsinochrome, a non-host selective toxin which contributes to virulence. However, the mechanisms involved in potential pathogen-host interactions occurring prior to the production of elsinochrome are unknown, yet the host-specificity observed among pathotypes suggests a reliance upon such mechanisms. In this study we have generated a whole genome sequencing project for E. fawcettii, producing an annotated draft assembly 26.01 Mb in size, with 10,080 predicted gene models and low (0.37%) coverage of transposable elements. A small proportion of the assembly showed evidence of AT-rich regions, potentially indicating genomic regions with increased plasticity. Using a variety of computational tools, we mined the E. fawcettii genome for potential virulence genes as candidates for future investigation. A total of 1,280 secreted proteins and 276 candidate effectors were predicted and compared to those of other necrotrophic (Botrytis cinerea, Parastagonospora nodorum, Pyrenophora tritici-repentis, Sclerotinia sclerotiorum and Zymoseptoria tritici), hemibiotrophic (Leptosphaeria maculans, Magnaporthe oryzae, Rhynchosporium commune and Verticillium dahliae) and biotrophic (Ustilago maydis) plant pathogens. Genomic and proteomic features of known fungal effectors were analysed and used to guide the prioritisation of 120 candidate effectors of E. fawcettii. Additionally, 378 carbohydrate-active enzymes were predicted and analysed for likely secretion and sequence similarity with known virulence genes. Furthermore, secondary metabolite prediction indicated nine additional genes potentially involved in the elsinochrome biosynthesis gene cluster than previously described. A further 21 secondary metabolite clusters were predicted, some with similarity to known toxin producing gene clusters. The candidate virulence genes predicted in this study provide a comprehensive resource for future experimental investigation into the pathogenesis of E. fawcettii.


Assuntos
Ascomicetos/genética , Citrus/microbiologia , Genoma Fúngico/genética , Anotação de Sequência Molecular , Ascomicetos/patogenicidade , Parede Celular/enzimologia , Mineração de Dados , Família Multigênica/genética , Metabolismo Secundário/genética
3.
Proc Natl Acad Sci U S A ; 117(7): 3492-3501, 2020 02 18.
Artigo em Inglês | MEDLINE | ID: mdl-32015115

RESUMO

Early detection and rapid response are crucial to avoid severe epidemics of exotic pathogens. However, most detection methods (molecular, serological, chemical) are logistically limited for large-scale survey of outbreaks due to intrinsic sampling issues and laboratory throughput. Evaluation of 10 canines trained for detection of a severe exotic phytobacterial arboreal pathogen, Candidatus Liberibacter asiaticus (CLas), demonstrated 0.9905 accuracy, 0.8579 sensitivity, and 0.9961 specificity. In a longitudinal study, cryptic CLas infections that remained subclinical visually were detected within 2 wk postinfection compared with 1 to 32 mo for qPCR. When allowed to interrogate a diverse range of in vivo pathogens infecting an international citrus pathogen collection, canines only reacted to Liberibacter pathogens of citrus and not to other bacterial, viral, or spiroplasma pathogens. Canines trained to detect CLas-infected citrus also alerted on CLas-infected tobacco and periwinkle, CLas-bearing psyllid insect vectors, and CLas cocultured with other bacteria but at CLas titers below the level of molecular detection. All of these observations suggest that canines can detect CLas directly rather than only host volatiles produced by the infection. Detection in orchards and residential properties was real time, ∼2 s per tree. Spatiotemporal epidemic simulations demonstrated that control of pathogen prevalence was possible and economically sustainable when canine detection was followed by intervention (i.e., culling infected individuals), whereas current methods of molecular (qPCR) and visual detection failed to contribute to the suppression of an exponential trajectory of infection.


Assuntos
Citrus/microbiologia , Cães/fisiologia , Doenças das Plantas/microbiologia , Rhizobiaceae/fisiologia , Olfato , Animais , Hemípteros/microbiologia , Hemípteros/fisiologia , Insetos Vetores/microbiologia , Insetos Vetores/fisiologia , Estudos Longitudinais , Rhizobiaceae/genética , Rhizobiaceae/isolamento & purificação
4.
PLoS Comput Biol ; 16(2): e1007570, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32027649

RESUMO

Diseases in humans, animals and plants remain an important challenge in our society. Effective control of invasive pathogens often requires coordinated concerted action of a large group of stakeholders. Both epidemiological and human behavioural factors influence the outcome of a disease control campaign. In mathematical models that are frequently used to guide such campaigns, human behaviour is often ill-represented, if at all. Existing models of human, animal and plant disease that do incorporate participation or compliance are often driven by pay-offs or direct observations of the disease state. It is however very well known that opinion is an important driving factor of human decision making. Here we consider the case study of Citrus Huanglongbing disease (HLB), which is an acute bacterial disease that threatens the sustainability of citrus production across the world. We show how by coupling an epidemiological model of this invasive disease with an opinion dynamics model we are able to answer the question: What makes or breaks the effectiveness of a disease control campaign? Frequent contact between stakeholders and advisors is shown to increase the probability of successful control. More surprisingly, we show that informing stakeholders about the effectiveness of control methods is of much greater importance than prematurely increasing their perceptions of the risk of infection. We discuss the overarching consequences of this finding and the effect on human as well as plant disease epidemics.


Assuntos
Citrus/microbiologia , Doenças das Plantas/prevenção & controle , Rhizobiaceae/patogenicidade , Surtos de Doenças , Modelos Teóricos , Doenças das Plantas/microbiologia , Estações do Ano
5.
Molecules ; 25(2)2020 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-31963827

RESUMO

To elucidate the role of fruit cuticle in fungal infection, changes in cuticle composition and morphology of 'Satsuma' mandarin during ambient (at 25 °C) storage and their role in Penicillium digitatum infection were investigated. Results showed that the epicuticular wax yield increased from 1.11 µg cm-2 to 4.21 µg cm-2 during storage for 20 days and then decreased to 1.35 µg cm-2 as storage time prolonged to 40 days. Intracuticular wax content of fruits stored for 20 days showed a peak value that was 1.7-fold higher than that of fruits stored for 40 days. The contents of cutin monomers of fruits showed a decreased trend during storage, while their proportions in the cutin stayed stable. Acids were identified as the most abundant components in epicuticular wax independently of the storage time, followed by alkanes and terpenoids. Terpenoids were found as the predominant components in intracuticular wax during the whole storage, followed by alkanes and acids. The flattened platelets crystals of fruits at harvest changed into small granule-like wax ones after 10 days of storage then gradually distributed across the surface of the fruits as stored for 40 days. Results of in vitro tests showed that mycelial growth of Penicillium digitatum could be promoted by epicuticular wax and conidial germination could be inhibited by cutin at different storage stages. These results shed new light on the chemical basis for cuticle involvement in fungal infection.


Assuntos
Citrus/anatomia & histologia , Citrus/microbiologia , Penicillium/fisiologia , Doenças das Plantas/microbiologia , Epiderme Vegetal/microbiologia , Citrus/efeitos dos fármacos , Frutas/anatomia & histologia , Lipídeos de Membrana/metabolismo , Esporos Fúngicos/crescimento & desenvolvimento , Ceras/química
6.
Artigo em Inglês | MEDLINE | ID: mdl-31945669

RESUMO

Citrus greening, also called Huanglongbing (HLB), is one of the most destructive citrus diseases worldwide. It is caused by the fastidious gram-negative α-proteobacteria bacterium Candidatus Liberibacter asiaticus (CLas) and vectored by the Asian citrus psyllid (ACP), Diaphorina citri. Currently, there is no cure for HLB, no compounds have been successful in controlling HLB, and no sustainable management practices have been established for the disease. Thus, searching for alternative citrus greening disease mitigation strategies is considered an urgent priority for a sustainable citrus industry. The aim of this study was to use compounds extracted from oak, Quercus hemisphaerica, and to assess the antibacterial effects of these against CLas-infected citrus plants. The application of aqueous oak leaf extracts showed substantial inhibitory effects against CLas in citrus plants and the activity of genes related to starch. Significant differences were also observed in plant phenotypic and physiological traits after treatments. Citrus plants treated with oak extracts displayed an increase in stomatal conductance, chlorophyll content and nutrient uptake concurrently with a reduction of CLas titer, when compared to citrus plants treated with just water. The information provided from this study suggests a new management treatment program to effectively deal with the HLB disease.


Assuntos
Citrus , Extratos Vegetais , Folhas de Planta , Quercus , Rhizobiaceae , Animais , Antibacterianos/farmacologia , Citrus/efeitos dos fármacos , Citrus/microbiologia , Hemípteros/microbiologia , Extratos Vegetais/farmacologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/microbiologia , Quercus/química , Rhizobiaceae/efeitos dos fármacos , Rhizobiaceae/fisiologia
7.
Food Microbiol ; 87: 103375, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31948616

RESUMO

This study investigated the biocontrol efficiency of Metschnikowia citriensis strain FL01 against Geotrichum citri-aurantii, and evaluated possible mechanisms. The results showed that M. citriensis could effectively control the development of sour rot, and significantly inhibit the mycelial growth and spore germination of G. citri-aurantii. The population dynamics results and Scanning electron microscopy (SEM) analysis indicated that M. citriensis could rapidly colonize wounds and tightly adhere to the surface of the wounds to compete with G. citri-aurantii for nutrition and space. M. citriensis also showed the biofilm formation action in vitro. The response of G. citri-aurantii to different components of M. citriensis culture showed that only the yeast cells but not the extracellular metabolites and the volatile organic compounds (VOCs) exhibited inhibitory effect on the growth of G. citri-aurantii. M. citriensis adhered to the hyphae of G. citri-aurantii loosely and sparsely, and the production of lytic enzymes ß-1, 3-glucanase (GLU) and Chitinase (CHI) could not be induced by G. citri-auranti. Iron affected the pulcherrimin pigment production and antagonism of M. citriensis indicating iron depletion as the most important antagonistic mechanism. Besides, M. citriensis also induced resistance of fruit against sour rot. These results suggested that M. citriensis could be used as the potential alternative of fungicides to control postharvest pathogens on citrus fruit.


Assuntos
Antibiose , Citrus/microbiologia , Geotrichum/crescimento & desenvolvimento , Metschnikowia/fisiologia , Doenças das Plantas/microbiologia , Frutas/microbiologia , Geotrichum/fisiologia , Metschnikowia/crescimento & desenvolvimento
8.
Sci Rep ; 10(1): 1389, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-31996756

RESUMO

Citrus black spot (CBS) caused by the fungus Phyllosticta citricarpa occurs in tropical and sub-tropical citrus production regions and affects all varieties of citrus. In Florida, the disease cycle is unique, having only the asexual spore. This work examines incidence and severity of CBS (hard spot symptoms) on fruit in two citrus groves during 2013-2014, 2014-2015 (Grove III) and 2015-2016 (Grove II) citrus seasons. Disease incidence and severity on fruit were analyzed based on citrus season, side of tree evaluated, height within the canopy, tree health, and tree age. Results indicate an increase in CBS incidence in Grove III between 2013-2014 and 2014-2015 seasons, with more infected or symptomatic fruit on the road side of the canopy and a higher incidence above 2 meters. Tree health status affected incidence but not severity and tree age had a significant effect on severity of CBS in Grove II. Analysis of weather data conducive for infection, between 2010 and 2017, indicated an average of 172 days per year (range: 104-261 days) when the temperature (15-35 °C) and relative humidity (RH ≥ 90% for 8 consecutive hours) were conducive for infection of fruit and an average of 98 days per year (range: 72-123 days) when the fruit were susceptible to infection.


Assuntos
Ascomicetos/isolamento & purificação , Citrus/microbiologia , Doenças das Plantas/microbiologia , Tempo (Meteorologia)
9.
Biochim Biophys Acta Gen Subj ; 1864(3): 129514, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31911239

RESUMO

BACKGROUND: Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker is maintained as an epiphyte on citrus leaves until entering the plant tissue. During epiphytic survival, bacteria may encounter low water availability that challenges the infection process. Proteomics analyses of Xcc under saline stress, mimicking the conditions found during epiphytic survival, showed increased abundance of a putative NAD(P)H dehydrogenase encoded by XAC2229. METHODS: Expression levels of XAC2229 and a Xcc mutant in XAC2229 were analyzed in salt and oxidative stress and during plant-pathogen interaction. An Escherichia coli expressing XAC2229 was obtained, and the role of this protein in oxidative stress resistance and in reactive oxygen species production was studied. Finally, Xac2229 protein was purified, spectrophotometric and cofactor analyses were done and enzymatic activities determined. RESULTS: XAC2229 was expressed under salt stress and during plant-pathogen interaction. ΔXAC2229 mutant showed less number of cankers and impaired epiphytic survival than the wild type strain. ΔXAC2229 survived less in the presence of H2O2 and produced more reactive oxygen species and thiobarbituric acid-reactive substances than the wild type strain. Similar results were observed for E. coli expressing XAC2229. Xac2229 is a FAD containing flavoprotein, displays diaphorase activity with an optimum at pH 6.0 and has quinone reductase activity using NADPH as an electron donor. CONCLUSIONS: A FAD containing flavoprotein from Xcc is a new NADPH quinone reductase required for bacterial virulence, particularly in Xcc epiphytic survival on citrus leaves. GENERAL SIGNIFICANCE: A novel protein involved in the worldwide disease citrus canker was characterized.


Assuntos
NAD(P)H Desidrogenase (Quinona)/metabolismo , Xanthomonas/enzimologia , Benzoquinonas/metabolismo , Citrus/metabolismo , Citrus/microbiologia , Peróxido de Hidrogênio/metabolismo , NAD(P)H Desidrogenase (Quinona)/genética , NADP/metabolismo , Estresse Oxidativo , Folhas de Planta/metabolismo , Estresse Salino/genética , Estresse Salino/fisiologia , Virulência , Xanthomonas/metabolismo , Xanthomonas/patogenicidade , Xanthomonas/fisiologia
10.
Mol Plant Microbe Interact ; 33(3): 519-527, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31973654

RESUMO

Xylella fastidiosa is a worldwide multihost pathogen that causes diseases in different crops. It is considered a new global threat and substantial efforts have been made in order to identify sources of resistance. Indeed, many genes have been associated with resistance to X. fastidiosa, but without functional validation. Here, we describe a C. reticulata gene homologous to the transcriptional factor RAP2.2 from Arabidopsis thaliana that increases resistance to citrus variegated chlorosis (CVC). This gene was previously detected in C. reticulata challenged with X. fastidiosa. Bioinformatics analysis together with subcellular localization and auto-activation assays indicated that RAP2.2 from C. reticulata (CrRAP2.2) is a transcriptional factor orthologous to AtRAP2.2. Thus, we used A. thaliana as a model host to evaluate the functional role of CrRAP2.2 in X. fastidiosa resistance. The inoculation of X. fastidiosa in the A. thaliana rap2.2 mutant resulted in a larger bacterial population, which was complemented by CrRAP2.2. In addition, symptoms of anthocyanin accumulation were higher in the mutant, whose phenotype was restored by CrRAP2.2, indicating that they have conserved functions in plant defense response. We therefore transformed C. sinensis with CrRAP2.2 and verified a positive correlation between CVC resistance and gene expression in transgenic lines. This is the first study using A. thaliana as model host that characterizes the function of a gene related to X. fastidiosa defense response and its application in genetic engineering to obtain citrus resistance to CVC.


Assuntos
Citrus/genética , Resistência à Doença/genética , Doenças das Plantas/genética , Fatores de Transcrição/genética , Xylella/patogenicidade , Arabidopsis , Proteínas de Arabidopsis , Citrus/microbiologia , Proteínas de Ligação a DNA , Doenças das Plantas/microbiologia
11.
J Basic Microbiol ; 60(1): 82-88, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31650621

RESUMO

Involvement of LaeA in various biological processes of filamentous fungi has been demonstrated. However, its role in Penicillium digitatum, the causal agent of citrus postharvest green mold, remains unclear. In this study, a ΔPdLaeA mutant was constructed using homologous recombination. The production of conidia by the ΔPdLaeA mutant was reduced by half compared with that of the wild-type strain. The sensitivity of the ΔPdLaeA mutant increased under alkaline conditions. The virulence assay revealed that PdLaeA was dispensable for the virulence of P. digitatum. Comparative transcriptome analysis revealed that the function loss of PdLaeA resulted in the reduced expression of several secondary metabolite gene clusters. In addition, expression of several key regulators of conidiation (BrlA, FlbA, FlbC, FlbD, and FluG) was also downregulated in the ΔPdLaeA mutant. In summary, the present work demonstrated that PdLaeA was involved in the regulation of SM biosynthesis, as well as the development and environmental adaptation of P. digitatum.


Assuntos
Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Penicillium/genética , Estresse Fisiológico/genética , Fatores de Transcrição/metabolismo , Citrus/microbiologia , Proteínas Fúngicas/genética , Perfilação da Expressão Gênica , Família Multigênica/genética , Penicillium/crescimento & desenvolvimento , Penicillium/metabolismo , Penicillium/fisiologia , Deleção de Sequência , Esporos Fúngicos/genética , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/metabolismo , Esporos Fúngicos/fisiologia , Fatores de Transcrição/genética , Virulência/genética
12.
Phytopathology ; 110(2): 287-296, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31713458

RESUMO

Citrus canker (CC), caused by the bacterial pathogen Xanthomonas citri subsp. citri, impacts citrus production in many areas of the world by reducing yields, degrading tree health, and severely blemishing the outer peels of fresh fruit. The relative susceptibility to CC among different species of Citrus varies from the highly susceptible lime (Citrus × aurantifolia), sweet orange (C. × sinensis), and grapefruit (C. × paradisi) to the much less susceptible calamondin (C. × microcarpa) and kumquat (C. japonica). This investigation compares the responses to infection with X. citri subsp. citri of these five genotypes with respect to phenylpropanoid compound profiles and relative increases or decreases of specific compounds postinoculation. In response to X. citri subsp. citri infection, all hosts possessed increased concentrations of phenylpropanoids in leaf tissue, whereas the similarly treated nonhost orange jessamine (Murraya paniculata) did not. Several of the tested genotypes exhibited notably increased production of fluorescent phenylpropanoids, including umbelliferone, herniarin, auraptene, scoparone, and others. The profiles of these compounds and their levels of production varied among the tested species yet all investigated Citrus genotypes exhibited increased concentrations of phenylpropanoids regardless of their degree of susceptibility to X. citri subsp. citri. Kumquat and calamondin, the tested genotypes least susceptible to X. citri subsp. citri, also exhibited the highest levels of the dihydrochalcone 3',5'-di-C-glucosyl phloretin, the aglycone portion of which, phloretin, is a known antibiotic, although levels of this compound were not affected by inoculation with X. citri subsp. citri.


Assuntos
Citrus , Interações Hospedeiro-Patógeno , Folhas de Planta , Xanthomonas , Citrus/microbiologia , Folhas de Planta/microbiologia , Xanthomonas/fisiologia
13.
Plant Dis ; 104(1): 239-245, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31710571

RESUMO

Citrus variegated chlorosis (CVC) disease, caused by the xylem-limited and insect-transmitted bacterium Xylella fastidiosa, has caused severe losses in orange production in Brazil. Disease control requires insecticide applications, tree removal, and pruning of symptomatic branches. Pruning success has been erratic, especially in areas of high disease incidence. In this work, in planta X. fastidiosa distribution and the effectiveness of severe pruning procedures for curing diseased adult trees were investigated. Most sampled upper parts of the trees contained X. fastidiosa, but at higher frequencies in symptomatic branches. Removal of all main branches (decapitation) was not effective and revealed a 20 to 30% incidence of latent infections. Trunk decapitation resulted in a higher number of healthy scions but killed 10 to 30% of the remaining trunks. Removal of all scion and grafting the newly sprouted shoots of 'Rangpur' lime (Citrus limonia Osbeck) or 'Cleopatra' (Citrus reshni Hort. ex Tan.) rootstocks with healthy buds allowed production of fast-growing and productive new scions that remained free from CVC for at least 2 years in four locations. With this method, highly affected trees do not need to be fully removed and the costs involved in this practice and in young tree acquisition and plantings are circumvented; therefore, it is a feasible option for less technically inclined small growers in Brazil.


Assuntos
Citrus sinensis , Citrus , Doenças das Plantas , Xylella , Brasil , Citrus/microbiologia , Doenças das Plantas/prevenção & controle , Xylella/fisiologia
14.
Phytopathology ; 110(2): 254-256, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31502518

RESUMO

Spiroplasma citri is a bacterium that causes stubborn disease of citrus and infects other crops, ornamentals, and weeds. It is transmitted by leafhoppers in a circulative manner. Due to limited sequence data on S. citri, the bacterium was isolated from naturally infected Chinese cabbage grown on a farm in Fresno County, CA. DNA from S. citri CC-2 was extracted from a pure culture in LD8 and subjected to PacBio sequencing. Four contigs were obtained with a single circular chromosome of 1,709,192 bp and three plasmids of 40,210, 39,313, and 2,921 bp in size. The genome developed herein extends the sequence database of S. citri and is the first whole-genome sequence record of S. citri from California.


Assuntos
Genoma Bacteriano , Doenças das Plantas , Spiroplasma citri , California , Citrus/microbiologia , Bases de Dados Genéticas , Genoma Bacteriano/genética , Doenças das Plantas/microbiologia , Spiroplasma citri/genética
15.
Mol Plant Microbe Interact ; 33(2): 135-137, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31577163

RESUMO

Elsinoë fawcettii and E. australis (phylum Ascomycota) are phytopathogenic fungi causing scab diseases on citrus plants. We report here the high-quality draft genome sequences and ab initio gene predictions of two E. fawcettii strains and one E. australis strain, which differ in their host range. This genome sequence information will provide valuable resources to underpin genomic attributes for determining host range through comparative genomic analyses of citrus scab fungi.


Assuntos
Ascomicetos , Citrus , Genoma Fúngico , Doenças das Plantas , Ascomicetos/genética , Citrus/microbiologia , Genoma Fúngico/genética , Especificidade de Hospedeiro , Doenças das Plantas/microbiologia
16.
Prep Biochem Biotechnol ; 50(3): 292-301, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31846377

RESUMO

Fungi are ecologically ubiquitous organisms on earth and regarded as one of the prolific sources of natural products. Fungal endophytes may provide essential prerequisite molecules to plant biochemical pathways which allow the efficient synthesis of primary and secondary metabolites. This study characterized the influences of various combinations of process parameters namely, carbohydrate, nitrogen, and phosphorus sources on citric acid (CA) production by the isolated fungal endophyte Aspergillus fumigatus P3I6 from Citrus microcarpa. Aspergillus fumigatus P3I6 had higher CA concentration of 9.2 (± 0.9) g L-1 and 9.0 (± 5.0 × 10-15) g L-1 when supplemented with sucrose and white refined sugar, respectively, than A. niger NRRL 599. Response Surface Methodology (RSM) had shown that A. fumigatus P3I6 produced the highest CA (23.8 g L-1) in Combination 4 (18.0% sucrose, 0.3 g L-1 ammonium sulfate, and 5.0 g L-1 dipotassium phosphate (K2HPO4)). Analysis of variance showed that when K2HPO4 concentrations were increased, CA content in fermentation media was significantly elevated. Hence, K2HPO4 was the most critical variable in the quadratic model (p < 0.05); however, sucrose concentration still has its role in production. Aside from using A. niger in most fermentation processes, this discovered fungal strain can be potentially used in biotechnological applications.


Assuntos
Aspergillus fumigatus/crescimento & desenvolvimento , Carboidratos/farmacologia , Ácido Cítrico/metabolismo , Citrus/microbiologia , Nitrogênio/farmacologia , Fósforo/farmacologia
17.
J Sci Food Agric ; 100(2): 744-754, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-31637724

RESUMO

BACKGROUND: The application of chemical fungicides is currently the main method for the control of postharvest decay of fruits and vegetables. However, public concern has been growing towards the effect of fungicides on food safety, human health and environmental pollution. Thus, interest in microbial biocontrol agent development has grown, such agents being considered both safe and environmentally friendly. Pseudomonas fluorescens is widely distributed in nature, and one of the most valuable biocontrol and plant growth-promoting rhizobacteria. In this study, the efficacy and the potential associated modes of action of P. fluorescens ZX against Penicillium italicum on oranges (Citrus sinensis Osbeck) were investigated. RESULTS: The application of P. fluorescens ZX significantly reduced blue mold lesion size and incidence in comparison to the control, where P. fluorescens ZX was effective when applied preventatively but not curatively. In dual cultures, treatment with cell-free autoclaved cultures or culture filtrate had a limited capacity to suppress P. italicum, while P. italicum was inhibited by bacterial fluid and bacterial suspension with living cells in vitro. The P. fluorescens ZX isolate displayed protease, but not chitinase, glucanase or cellulose, activity, and produced siderophores and volatile organic compounds with antifungal abilities. Competition tests showed P. fluorescens ZX could use fructose, sucrose, aspartic acid, threonine, serine, glycine, valine, lysine and proline better than P. italicum. Furthermore, an effective biofilm that peaked after a 24-hour incubation at 30 °C was formed by the P. fluorescens ZX isolate. Light microscopy and scanning electron microscopy observations indicate the P. fluorescens ZX isolate could not undergo direct parasitism or hyperparasitism. CONCLUSIONS: Competition for nutrients and niches, biofilm formation, inhibition of spore germination and mycelial growth, and production of inhibitory metabolites may play important roles in P. fluorescens ZX antagonism of P. italicum. © 2019 Society of Chemical Industry.


Assuntos
Antibiose , Citrus/microbiologia , Pseudomonas fluorescens/fisiologia , Biofilmes , Frutas/microbiologia , Micélio/crescimento & desenvolvimento , Micélio/metabolismo , Penicillium/crescimento & desenvolvimento , Penicillium/metabolismo , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle
18.
J Appl Microbiol ; 128(5): 1400-1413, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31841255

RESUMO

AIMS: This study aimed at isolating endophytic fungi from Citrus limon (L.) possessing antioxidative and genoprotective potential. METHODS AND RESULTS: Endophytic fungi were screened for antioxidant activity using 2,2-diphenyl,1-picryl hydrazyl radical scavenging assay and maximum activity (79·70%) was exhibited by culture MP1 identified to be Penicillium oxalicum on the basis of morphological and molecular characteristics. The ethyl acetate extract of MP1 was subjected to silica column chromatography followed by LH 20 column chromatography for purification of active metabolites. The partially purified active fraction of P. oxalicum MP1 possessed good antioxidant activity as detected using various assays. It also exhibited a strong DNA damage protection potential on pUC19 plasmid DNA treated with Fenton reagent. On exposure to active fraction of MP1 significant reduction (P < 0·05) in nuclear deformities (like nuclear buds, micronuclei, nuclear ridges and binucleated cells) was observed in human lymphocytes pretreated with a toxic concentration of H2 O2 . In vivo genoprotectivity studies were conducted in fresh water fish Channa punctatus pretreated with a damaging compound 4-nonyl phenol. The active fraction of P. oxalicum MP1 caused a reduction of 94·7 and 66·60% in micronuclei and aberrant cell formation, respectively. A significant reduction (P < 0·05) in tail length and tail DNA parameters was also observed in comet assay. CONCLUSION: The endophytic P. oxalicum isolated in this study has the potential to produce metabolites possessing antioxidant and genoprotective activities. SIGNIFICANCE AND IMPACT OF THE STUDY: The isolated culture can be exploited in the field of therapeutics by virtue of its in vitro and in vivo genoprotective potential.


Assuntos
Antimutagênicos/metabolismo , Antioxidantes/metabolismo , Citrus/microbiologia , Penicillium/isolamento & purificação , Penicillium/metabolismo , Animais , Antimutagênicos/farmacologia , Antioxidantes/farmacologia , Dano ao DNA/efeitos dos fármacos , Endófitos , Humanos , Penicillium/classificação
19.
BMC Genomics ; 20(1): 969, 2019 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-31829190

RESUMO

BACKGROUND: Citrus blight is a very important progressive decline disease of commercial citrus. The etiology is unknown, although the disease can be transmitted by root grafts, suggesting a viral etiology. Diagnosis is made by demonstrating physical blockage of xylem cells that prevents the movement of water. This test was used to identify symptomatic trees from four commercial groves in Florida. Total RNA extracts of phloem-enriched scaffold root tissues were prepared from seven trees that failed to take up water and from one healthy tree. These RNA extracts were used for transcriptomic analyses using paired end RNA-Seq from an Illumina 2500 system. The expression of transcripts annotated as polyprotein of citrus endogenous pararetrovirus were estimated by both RT-qPCR and RNA-Seq. RESULTS: Transcripts from seven RNA-Seq libraries from trees affected by citrus blight were compared to a control tree. 129-148 million RNA fragments (two paired-end reads/fragment) were generated per library and were mapped to the sweet orange reference genome. In response to citrus blight stress, genes encoding aquaporins, proteins with water channel activity and several cellulose synthase genes were down-regulated, whereas genes involved in lignin and glucosinolate biosynthesis were up-regulated. Transcripts encoding proteins in pathways of carbohydrate metabolism, nucleotide synthesis, signaling, hormone metabolism, secondary metabolism, transport, and biotic stress pathways were overwhelmingly down regulated in all libraries. CONCLUSION: Reduced water intake and xylem plugging were observed in the trees tested and the changes in their transcriptome were analyzed. Plants adapted to reduced water flow by regulating primary and secondary metabolism, nuclear transport and hormone associated pathways. The patterns of energy generation, transcription, translation and protein degradation were consistent with irreversible decline. The down regulation of cellulose synthase transcripts and up regulation of transcripts related to lignin production likely lead to an imbalance in the pathways leading to wood formation, and may lead to the blockage of the xylem vessels seen as the cardinal symptom of citrus blight. Transcripts of a pararetrovirus were elevated in the transcriptome of roots used in this study.


Assuntos
Citrus/fisiologia , Perfilação da Expressão Gênica/métodos , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Citrus/microbiologia , Regulação da Expressão Gênica de Plantas , Biblioteca Gênica , Doenças das Plantas/genética , Raízes de Plantas/microbiologia , Raízes de Plantas/fisiologia , Metabolismo Secundário , Análise de Sequência de RNA , Água/metabolismo , Xilema/metabolismo
20.
World J Microbiol Biotechnol ; 36(1): 7, 2019 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-31845300

RESUMO

Orange juice is a major agricultural product, and oranges are among the most widely sold fresh fruits in several countries. Citrus black spot (CBS), caused by the fungus Phyllosticta citricarpa, affects almost every species of citrus, causing a esthetic depreciation of the fruit and fruit drop, with consequent financial loss for its in natura commercialization. Fungicides are the major control measure for CBS, but have limited efficiency and high cost, and give rise to fungal strains resistant to these products. This work assayed the production of antifungal compounds by Bacillus spp. isolates and tested the potential of two isolates from B. subtilis (ACB-AP3 and ACB-83) for controlling CBS under field conditions with two previously untested orange varieties. A total of 15 isolates produced cell-free, thermostable or volatile compounds effective in suppressing P. citricarpa growth in vitro. It was possible to detect the production of two antibiotics (iturin and surfactin) by B. subtilis ACB-83. Additionally, B. subtilis isolates ACB-AP3 and ACB-83 displayed significant effects in controlling the disease under field conditions.


Assuntos
Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Ascomicetos/efeitos dos fármacos , Bacillus/isolamento & purificação , Bacillus/metabolismo , Citrus/microbiologia , Ascomicetos/crescimento & desenvolvimento , Ascomicetos/patogenicidade , Frutas/microbiologia , Fungicidas Industriais/metabolismo , Fungicidas Industriais/farmacologia , Doenças das Plantas/microbiologia , Esporos Fúngicos
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