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1.
Int J Food Microbiol ; 335: 108889, 2020 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-33007604

RESUMO

The safety and quality of cereal grain supplies are adversely impacted by microbiological contamination, with novel interventions required to maximise whole grains safety and stability. The microbiological contaminants of wheat grains and the efficacy of Atmospheric Cold Plasma (ACP) for potential to control these risks were investigated. The evaluations were performed using a contained reactor dielectric barrier discharge (DBD) system; samples were treated for 0-20 min using direct and indirect plasma exposure. Amplicon-based metagenomic analysis using bacterial 16S rRNA gene and fungal 18S rRNA gene with internal transcribed spacer (ITS) region was performed to characterize the change in microbial community composition in response to ACP treatment. The antimicrobial efficacy of ACP against a range of bacterial and fungal contaminants of wheat, was assessed to include individual isolates from grains as challenge pathogens. ACP influenced wheat microbiome composition, with a higher microbial diversity as well as abundance found on the untreated control grain samples. Culture and genomic approaches revealed different trends for mycoflora detection and control. A challenge study demonstrated that using direct mode of plasma exposure with 20 min of treatment significantly reduced the concentration of all pathogens. Overall, reduction levels for B. atrophaeus vegetative cells were higher than for all fungal species tested, whereas B. atrophaeus spores were the most resistant to ACP among all microorganisms tested. Of note, repeating sub-lethal plasma treatment did not induce resistance to ACP in either B. atrophaeus or A. flavus spores. ACP process control could be tailored to address diverse microbiological risks for grain stability and safety.


Assuntos
Anti-Infecciosos/farmacologia , Resistência Microbiana a Medicamentos , Microbiota/efeitos dos fármacos , Gases em Plasma/farmacologia , Triticum/microbiologia , Bactérias/classificação , Bactérias/efeitos dos fármacos , Bactérias/genética , Bactérias/isolamento & purificação , Grão Comestível/microbiologia , Fungos/classificação , Fungos/efeitos dos fármacos , Fungos/genética , Fungos/isolamento & purificação , RNA Ribossômico/genética , Especificidade da Espécie
2.
PLoS One ; 15(9): e0238724, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32956369

RESUMO

Wheat blast caused by Magnaporthe oryzae pathotype Triticum (MoT) is a threat to wheat production especially in the warmer-humid environments. In Zambia, wheat blast symptoms were observed for the first time on wheat (Triticum aestivum L.) grown in experimental plots and five farmers' fields in Mpika district of Muchinga Province during the 2017-18 rainy season. Infected plants showed the typical wheat blast symptoms with the spike becoming partially or completely bleached with the blackening of the rachis in a short span of time. Incidence of blast symptoms on nearly all wheat heads was high and ranged from 50 to 100%. Examination of diseased plant leaves showed the presence of elliptical, grayish to tan necrotic lesions with dark borders on the leaf often mixed with other foliar diseases. A study was conducted to isolate and identify the causal pathogen(s) using classical and molecular methods and determine the pathogenicity of the detected disease causal agent. Morphobiometrical determination of causal pathogen revealed conidia with characteristic pear shaped 2-septate hyaline spores associated with M. oryzae species. Preliminary polymerase chain reaction screening of six isolates obtained from wheat blast infected samples with diagnostic primers (MoT3F/R) was conducted at ZARI, Zambia, and subsequent analysis of two isolates with MoT3F/R and C17F/R was performed at USDA-ARS, USA. Both experiments confirmed that MoT is the causal agent of wheat blast in Zambia. Further, pathogenicity tests performed with pure culture isolates from samples WS4 and WS5 produced typical blast symptoms on all the six inoculated wheat genotypes. Results of this study indicate that MoT is causing wheat blast in rain-fed wheat grown in Zambia, thus making it the first report of MoT in Zambia and Africa. This inter-continental movement of the pathogen (disease) has serious implication for wheat production and trade that needs to be urgently addressed.


Assuntos
Magnaporthe/isolamento & purificação , Magnaporthe/fisiologia , Doenças das Plantas/microbiologia , Triticum/microbiologia , Magnaporthe/patogenicidade , Esporos Fúngicos/isolamento & purificação , Esporos Fúngicos/fisiologia , Zâmbia
3.
PLoS One ; 15(9): e0237460, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32911506

RESUMO

Infection of wheat by Fusarium species can lead to Fusarium Head Blight (FHB) and mycotoxin contamination, thereby reducing food quality and food safety, and leading to economic losses. Agronomic management through the implementation of various pre-harvest measures can reduce the probability of Fusarium spp. infection in the wheat field. To design interventions that could stimulate wheat farmers to (further) improve their agronomic management to reduce FHB, it is key to understand farmers' behaviour towards adapting their management. The aim of this paper was to understand the intention, underlying behavioural constructs, and beliefs of Dutch wheat farmers to adapt their agronomic management to reduce FHB and mycotoxin contamination in wheat, applying the Theory of Planned Behaviour (TPB). Data were collected from 100 Dutch wheat farmers via a questionnaire. The standard TPB analysis was extended with an assessment of the robustness of the belief results to account for the statistical validity of the analysis on TPB beliefs (i.e. to address the so-called expectancy-value muddle). Forty-six percent of the farmers had a positive intention to change their management in the next 5 years. The two behavioural constructs significantly related to this intention were attitude and social norm, whereas association with the perceived behavioural control construct was insignificant indicating that farmers did not perceive any barriers to change their behaviour. Relevant attitudinal beliefs indicated specific attributes of wheat, namely yield, quality and safety (lower mycotoxin contamination). This indicates that strengthening these beliefs-by demonstrating that a change in management will result in a higher yield and quality and lower mycotoxin levels-will result in a stronger attitude and, subsequently, a higher intention to change management. Interventions to strengthen these beliefs should preferably go by the most important referents for social norms, which were the buyers and the farmer cooperatives in this study.


Assuntos
Produção Agrícola/métodos , Fusarium/isolamento & purificação , Doenças das Plantas/prevenção & controle , Triticum/microbiologia , Agricultura/métodos , Controle Comportamental , Fazendeiros , Qualidade dos Alimentos , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Intenção , Doenças das Plantas/microbiologia , Normas Sociais , Inquéritos e Questionários
4.
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
5.
Sheng Wu Gong Cheng Xue Bao ; 36(7): 1356-1364, 2020 Jul 25.
Artigo em Chinês | MEDLINE | ID: mdl-32748593

RESUMO

Salinity is the most important factor for the growth of crops. It is an effective method to alleviate the toxic effect caused by salt stress using saline-alkali-tolerant and growth-promoting bacteria in agriculture. Seven salt-tolerant bacteria were screened from saline-alkali soil, and the abilities of EPS production, alkalinity reduction and IAA production of the selected strains were investigated. A dominant strain DB01 was evaluated. The abilities of EPS production, alkalinity reduction and IAA production of strain DB01 were 0.21 g/g, 8.7% and 8.97 mg/L, respectively. The isolate was identified as Halomonas aquamarina by partial sequencing analysis of its 16S rRNA genes, and had the ability to inhibit the growth of Fusarium oxysporum f. sp., Alternaria solani, Phytophthora sojae and Rhizoctonia cerealis. It also could promote root length and germination rate of wheat seedlings under salt stress. Halomonas aquamarina can provide theoretical basis for the development of soil microbial resources and the application in saline-alkali soil improvement.


Assuntos
Álcalis , Raízes de Plantas , Tolerância ao Sal , Microbiologia do Solo , Álcalis/metabolismo , Bactérias/efeitos dos fármacos , Bactérias/genética , Halomonas/genética , Raízes de Plantas/microbiologia , RNA Ribossômico 16S/genética , Tolerância ao Sal/genética , Plântula/crescimento & desenvolvimento , Plântula/microbiologia , Solo/química , Triticum/microbiologia
6.
Int J Food Microbiol ; 334: 108808, 2020 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-32835995

RESUMO

Heat-stable mycotoxins are widely detected in flour and produced by Aspergillus spp., Fusarium spp. and Penicillium spp. Forty different flours purchased in Italy are used to assess potential risk factors via a systematically screening of a number of variables: the type of flour, organic, whole and white wheat, types of packaging (paper, plastic and weight). Fungal recovery and co-occurrence of specific mycotoxins was also assessed. The results showed that flour originated from fruits had a significant higher recovery of fungi, while seed/pseudocereals had the highest mycotoxins detection. Flours originating from organic agriculture are more prone to higher fungal recovery and mycotoxins detection when compared with not-organic flours. Packaging is also important: packaging weighting less than 376 g supports significantly more fungal recovery and the plastic packages was observed to retain more fungi and mycotoxins detection when compared with paper. Recovery measured as Log (CFU/g) of fungal genera is not directly proportional to the amount of mycotoxins. Finally, linear regression and mixed logit regression models show that the mean level of aflatoxins B1 (ng/g on the logarithmic scale) reduces by 0.485 when moving from an organic to a non-organic flour, while a significant increase of 0.369 when moving from paper to a plastic packaging.


Assuntos
Farinha/análise , Farinha/microbiologia , Contaminação de Alimentos/análise , Fungos/isolamento & purificação , Micotoxinas/análise , Embalagem de Alimentos , Fungos/classificação , Fungos/metabolismo , Itália , Agricultura Orgânica , Triticum/microbiologia
7.
PLoS One ; 15(8): e0233665, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32804955

RESUMO

Oligomycins are macrolide antibiotics, produced by Streptomyces spp. that show antagonistic effects against several microorganisms such as bacteria, fungi, nematodes and the oomycete Plasmopara viticola. Conidiogenesis, germination of conidia and formation of appressoria are determining factors pertaining to pathogenicity and successful diseases cycles of filamentous fungal phytopathogens. The goal of this research was to evaluate the in vitro suppressive effects of two oligomycins, oligomycin B and F along with a commercial fungicide Nativo® 75WG on hyphal growth, conidiogenesis, conidial germination, and appressorial formation of the wheat blast fungus, Magnaporthe oryzae Triticum (MoT) pathotype. We also determined the efficacy of these two oligomycins and the fungicide product in vivo in suppressing wheat blast with a detached leaf assay. Both oligomycins suppressed the growth of MoT mycelium in a dose dependent manner. Between the two natural products, oligomycin F provided higher inhibition of MoT hyphal growth compared to oligomycin B with a minimum inhibitory concentration of 0.005 and 0.05 µg/disk, respectively. The application of the compounds completely halted conidial formation of the MoT mycelium in agar medium. Further bioassays showed that these compounds significantly inhibited MoT conidia germination and induced lysis. The compounds also caused abnormal germ tube formation and suppressed appressorial formation of germinated spores. Interestingly, the application of these macrolides significantly inhibited wheat blast on detached leaves of wheat. This is the first report on the inhibition of mycelial growth, conidiogenesis, germination of conidia, deleterious morphological changes in germinated conidia, and suppression of blast disease of wheat by oligomycins from Streptomyces spp. Further study is needed to unravel the precise mode of action of these natural compounds and consider them as biopesticides for controlling wheat blast.


Assuntos
Magnaporthe/efeitos dos fármacos , Magnaporthe/patogenicidade , Oligomicinas/farmacologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Triticum/microbiologia , Agentes de Controle Biológico/farmacologia , Grão Comestível/microbiologia , Microbiologia de Alimentos , Fungicidas Industriais/farmacologia , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Magnaporthe/crescimento & desenvolvimento , Micélio/efeitos dos fármacos , Micélio/crescimento & desenvolvimento , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/crescimento & desenvolvimento
8.
Int J Food Microbiol ; 333: 108777, 2020 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-32745828

RESUMO

This study aimed to model the aflatoxin B1 (AFB1) production by A. flavus in wheat grains during malting for craft beer. A total of sixty-four different combinations of grains steeping degree (ST; 41, 43, 45 and 47%), temperature (13, 15, 17 and 19 °C) and time of germination (48, 72, 96 and 120 h), comprising the range of malting conditions that allow the production of quality malt, were assayed. AFB1 was produced in a range of 15.78 ± 3.54 µg/kg (41% ST, 13 °C for 48 h) to 284.66 ± 44.34 µg/kg (47% ST, 19 °C for 120 h). The regression model showing an acceptable fit to the experimental data (adjusted R2 0.84) for AFB1 as a function of grains steeping degree, temperature and time of germination. Results showed that AFB1 levels in wheat malt increase with increase of the temperature or time of germination. Within the range of tested malting conditions, no significant effects were observed for steeping degree on AFB1 levels in wheat malt. The generated model is useful to estimate the AFB1 levels in wheat malt. Findings highlight overall that if wheat grains are contaminated with A. flavus, AFB1 might be produced in malt in levels above the limits set by regulatory agencies, regardless the steeping conditions used.


Assuntos
Aflatoxina B1/biossíntese , Aspergillus flavus/metabolismo , Cerveja/microbiologia , Modelos Biológicos , Triticum/microbiologia , Cerveja/análise , Fermentação/fisiologia , Germinação/fisiologia , Sementes/microbiologia , Temperatura
9.
Appl Environ Microbiol ; 86(17)2020 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-32631868

RESUMO

A large number of strains in the Rhizobium radiobacter species complex (biovar 1 Agrobacterium) have been known as causative pathogens for crown gall and hairy root diseases. Strains within this complex were also found as endophytes in many plant species with no symptoms. The aim of this study was to reveal the endophyte variation of this complex and how these endophytic strains differ from pathogenic strains. In this study, we devised a simple but effective screening method by exploiting the high resolution power of mass spectrometry. We screened endophyte isolates from young wheat and barley plants, which are resistant to the diseases, and identified seven isolates from wheat as members of the R. radiobacter species complex. Through further analyses, we assigned five strains to the genomovar (genomic group) G1 and two strains to G7 in R. radiobacter Notably, these two genomovar groups harbor many known pathogenic strains. In fact, the two G7 endophyte strains showed pathogenicity on tobacco, as well as the virulence prerequisites, including a 200-kbp Ri plasmid. All five G1 strains possessed a 500-kbp plasmid, which is present in well-known crown gall pathogens. These data strongly suggest that healthy wheat plants are reservoirs for pathogenic strains of R. radiobacter IMPORTANCE Crown gall and hairy root diseases exhibit very wide host-plant ranges that cover gymnosperm and dicot plants. The Rhizobium radiobacter species complex harbors causative agents of the two diseases. Recently, endophyte isolates from many plant species have been assigned to this species complex. We isolated seven endophyte strains belonging to the species complex from wheat plants and revealed their genomovar affiliations and plasmid profile. The significance of this study is the finding of the genomovar correlation between the endophytes and the known pathogens, the presence of a virulence ability in two of the seven endophyte strains, and the high ratio of the pathogenic strains in the endophyte strains. This study therefore provides convincing evidence that could unravel the mechanism that maintains pathogenic agents of this species and sporadically delivers them to susceptible plants.


Assuntos
Agrobacterium tumefaciens/fisiologia , Reservatórios de Doenças/microbiologia , Endófitos/fisiologia , Hordeum/microbiologia , Doenças das Plantas/microbiologia , Plasmídeos/isolamento & purificação , Triticum/microbiologia
10.
Plant Mol Biol ; 104(1-2): 113-136, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32627097

RESUMO

KEY MESSAGE: Present study revealed a complex relationship among histone H3 methylation (examined using H3K4/K27me3 marks), cytosine DNA methylation and differential gene expression during Lr28 mediated leaf rust resistance in wheat. During the present study, genome-wide histone modifications were examined in a pair of near isogenic lines (NILs) (with and without Lr28 in the background of cv. HD2329). The two histone marks used included H3K4me3 (an activation mark) and H3K27me3 (a repression mark). The results were compared with levels of expression (using RNA-seq) and DNA methylation (MeDIP) data obtained using the same pair of NILs. Some of the salient features of the present study include the following: (i) large scale differential binding sites (DBS) were available for only H3K4me3 in the susceptible cultivar, but for both H3K4me3 and H3K27me3 in its resistant NIL; (ii) DBSs for H3K27me3 mark were more abundant (> 80%) in intergenic regions, whereas DBSs for H3K4me3 were distributed in all genomic regions including exons, introns, intergenic, TTS (transcription termination sites) and promoters; (iii) fourteen (14) genes associated with DBSs showed co-localization for both the marks; (iv) only a small fraction (7% for H3K4me3 and 12% for H3K27me3) of genes associated with DBSs matched with the levels of gene expression inferred from RNA-seq data; (v) validation studies using qRT-PCR were conducted on 26 selected representative genes; results for only 11 genes could be validated. The proteins encoded by important genes involved in promoting infection included domains generally carried by R gene proteins such as Mlo like protein, protein kinases and purple acid phosphatase. Similarly, proteins encoded by genes involved in resistance included those carrying domains for lectin kinase, R gene, aspartyl protease, etc. Overall, the results suggest a very complex network of downstream genes that are expressed during compatible and incompatible interactions; some of the genes identified during the present study may be used in future validation studies involving RNAi/overexpression approaches.


Assuntos
Basidiomycota/metabolismo , Resistência à Doença/genética , Genes de Plantas/genética , Genoma de Planta/genética , Histonas/genética , Doenças das Plantas/genética , Triticum/genética , Triticum/metabolismo , Imunoprecipitação da Cromatina , Metilação de DNA , Regulação da Expressão Gênica de Plantas , Ligação Genética , Histonas/metabolismo , Anotação de Sequência Molecular , Doenças das Plantas/microbiologia , Folhas de Planta/genética , Folhas de Planta/microbiologia , Regiões Promotoras Genéticas , Reprodutibilidade dos Testes , Alinhamento de Sequência , Análise de Sequência , Análise de Sequência de RNA , Transcrição Genética , Triticum/microbiologia
11.
Food Chem ; 332: 127288, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32619935

RESUMO

This study examined the effect of green tea extract at 10 (GWG1%) and 50 (GWG5%) g/L as the steeping solution on the chemical, nutritional, and microbial quality of wheat grain during 14 days of germination. Fat, dry matter, and ash contents in the control was higher than GWG treatments due to the faster growth of control germs. Moisture, phenolic compounds, thiamin, niacin, and tocopherols decreased, whereas, fat, dry matter, carbohydrate, protein, crude fiber, ash, folic acid, Ca, Mg, Fe, Mn, and Zn increased significantly in all samples during germination. GWG5% showed the highest values in total phenols, vitamins, minerals, and carbohydrate, followed by GWG1% and then the control. No significant differences in protein and crude fiber content were detected among treatments. GWG decreased the growth of total bacterial, yeast, and mold in germinated seeds. Overall, GWG improved the microbial and nutritional quality of wheat germ during 14 days of germination.


Assuntos
Germinação/efeitos dos fármacos , Valor Nutritivo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Chá/química , Triticum/metabolismo , Triticum/microbiologia , Triticum/efeitos dos fármacos
12.
PLoS One ; 15(6): e0235482, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32603342

RESUMO

Fusarium head blight (FHB) is one of the most devastating fungal diseases affecting grain crops and Fusarium graminearum is the most aggressive causal species. Several evidences shown that stomatal closure is involved in the first line of defence against plant pathogens. However, there is very little evidence to show that photosynthetic parameters change in inoculated plants. The aim of the present study was to study the role of stomatal regulation in wheat after F. graminearum inoculation and explore its possible involvement in FHB resistance. RT-qPCR revealed that genes involved in stomatal regulation are induced in the resistant Sumai3 cultivar but not in the susceptible Rebelde cultivar. Seven genes involved in the positive regulation of stomatal closure were up-regulated in Sumai3, but it is most likely, that two genes, TaBG and TaCYP450, involved in the negative regulation of stomatal closure, were strongly induced, suggesting that FHB response is linked to cross-talk between the genes promoting and inhibiting stomatal closure. Increasing temperature of spikes in the wheat genotypes and a decrease in photosynthetic efficiency in Rebelde but not in Sumai3, were observed, confirming the hypothesis that photosynthetic parameters are related to FHB resistance.


Assuntos
Resistência à Doença/genética , Fusariose/imunologia , Fotossíntese , Estômatos de Plantas , Triticum/imunologia , Fusarium/imunologia , Fusarium/patogenicidade , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Fotossíntese/genética , Fotossíntese/imunologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Imunidade Vegetal/genética , Estômatos de Plantas/genética , Estômatos de Plantas/imunologia , Estômatos de Plantas/fisiologia , Triticum/genética , Triticum/microbiologia
13.
PLoS Genet ; 16(7): e1008713, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32658889

RESUMO

Thaumatin-like proteins (TLPs), which are defined as pathogenesis-related protein family 5 (PR5) members, are common plant proteins involved in defense responses and confer antifungal activity against many plant pathogens. Our earlier studies have reported that the TaTLP1 gene was isolated from wheat and proved to be involved in wheat defense in response to leaf rust attack. The present study aims to identify the interacting proteins of TaTLP1 and characterize the role of the interaction between wheat and Puccinia triticina (Pt). Pull-down experiments designed to isolate the molecular target of TaTLP1 in tobacco resulted in the identification of TaPR1, a pathogenesis-related protein of family 1, and the interaction between TaTLP1 and TaPR1 was confirmed by yeast two-hybrid experiments (Y2H), bimolecular fluorescence complementation (BiFC), and co-immunoprecipitation (Co-IP). In vitro, TaTLP1 and TaPR1 together increased antifungal activity against Pt. In vivo, the disease resistance phenotype, histological observations of fungal growth and host responses, and accumulation of H2O2 in TaTLP1-TaPR1 in co-silenced plants indicated that co-silencing significantly enhanced wheat susceptibility compared to single knockdown TaTLP1 or TaPR1 plants. The accumulation of reactive oxygen species (ROS) was significantly reduced in co-silenced plants compared to controls during Pt infection, which suggested that the TaTLP1-TaPR1 interaction positively modulates wheat resistance to Pt in an ROS-dependent manner. Our findings provide new insights for understanding the roles of two different PRs, TaTLP1 and TaPR1, in wheat resistance to leaf rust.


Assuntos
Antígenos de Plantas/genética , Resistência à Doença/genética , Doenças das Plantas/genética , Proteínas de Plantas/genética , Triticum/genética , Basidiomycota/genética , Basidiomycota/patogenicidade , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Espécies Reativas de Oxigênio/metabolismo , Triticum/crescimento & desenvolvimento , Triticum/microbiologia
14.
Food Chem ; 332: 127420, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32622191

RESUMO

The outbreaks of Cronobacter sakazakii, Salmonella spp, and Bacillus cereus in powdered foods have been increasing in worldwide. However, an effective method to pasteurize powdered foods before consumption remains lacking. A prototype Intense Pulsed Light (IPL) system was developed to disinfect powdered foods under different IPL and environmental conditions. Synergistic effect of IPL and TiO2 photocatalysis on microbial inactivation was studied. The results show that high energy intensity of each pulse, high peak intensity, and short pulsed duration contributed to a high microbe inactivation. With TiO2 photocatalysis, one additional log10 reduction was achieved, bringing the total log reduction to 4.71 ± 0.07 (C. sakazakii), 3.49 ± 0.01 (E. faecium), and 2.52 ± 0.10 (B. cereus) in non-fat dry milk, and 5.42 ± 0.10 (C. sakazakii), 4.95 ± 0.24 (E. faecium), 2.80 ± 0.23 (B. cereus) in wheat flour. IPL treatment combined with the TiO2 photocatalysis exhibits a strong potential to reduce the energy consumption in improving the safety of powdered foods.


Assuntos
Cronobacter sakazakii/efeitos da radiação , Cronobacter/efeitos da radiação , Farinha/microbiologia , Conservação de Alimentos/métodos , Viabilidade Microbiana/efeitos da radiação , Leite/microbiologia , Triticum/microbiologia , Animais , Bacillus cereus/crescimento & desenvolvimento , Bacillus cereus/efeitos da radiação , Cronobacter/crescimento & desenvolvimento , Microbiologia de Alimentos , Conservação de Alimentos/instrumentação , Luz , Pós/química , Salmonella/crescimento & desenvolvimento , Salmonella/efeitos da radiação
15.
Mol Plant Microbe Interact ; 33(7): 888-901, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32484730

RESUMO

The plant-pathogenic fungus Fusarium graminearum, causal agent of Fusarium head blight (FHB) disease on small grain cereals, produces toxic trichothecenes that require facilitated export for full virulence. Two potential modes of mycotoxin transport are membrane-bound transporters, which move toxins across cellular membranes, and N-ethylmaleimide-sensitive factor attachment receptor (SNARE)-mediated vesicular transport, by which toxins may be packaged as cargo in vesicles bound for organelles or the plasma membrane. In this study, we show that deletion of a gene (Sso2) for a subapically localized t-SNARE protein results in growth alteration, increased sensitivity to xenobiotics, altered gene expression profiles, and reduced deoxynivalenol (DON) accumulation in vitro and in planta as well as reduced FHB symptoms on wheat. A double deletion mutant generated by crossing the ∆sso2 deletion mutant with an ATP-binding cassette transporter deletion mutant (∆abc1) resulted in an additive reduction in DON accumulation and almost complete loss of FHB symptoms in planta. These results suggest an important role of Sso2-mediated subapical exocytosis in FHB progression and xenobiotic defense and are the first report of an additive reduction in F. graminearum DON accumulation upon deletion of two distinct modes of cellular export. This research provides useful information which may aid in formulating novel management plans of FHB or other destructive plant diseases.


Assuntos
Fusarium/patogenicidade , Doenças das Plantas/microbiologia , Proteínas SNARE/genética , Tricotecenos/metabolismo , Fusarium/genética , Deleção de Genes , Genes Fúngicos , Triticum/microbiologia , Virulência
16.
PLoS Pathog ; 16(6): e1008652, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32574207

RESUMO

Plants trigger immune responses upon recognition of fungal cell wall chitin, followed by the release of various antimicrobials, including chitinase enzymes that hydrolyze chitin. In turn, many fungal pathogens secrete LysM effectors that prevent chitin recognition by the host through scavenging of chitin oligomers. We previously showed that intrachain LysM dimerization of the Cladosporium fulvum effector Ecp6 confers an ultrahigh-affinity binding groove that competitively sequesters chitin oligomers from host immune receptors. Additionally, particular LysM effectors are found to protect fungal hyphae against chitinase hydrolysis during host colonization. However, the molecular basis for the protection of fungal cell walls against hydrolysis remained unclear. Here, we determined a crystal structure of the single LysM domain-containing effector Mg1LysM of the wheat pathogen Zymoseptoria tritici and reveal that Mg1LysM is involved in the formation of two kinds of dimers; a chitin-dependent dimer as well as a chitin-independent homodimer. In this manner, Mg1LysM gains the capacity to form a supramolecular structure by chitin-induced oligomerization of chitin-independent Mg1LysM homodimers, a property that confers protection to fungal cell walls against host chitinases.


Assuntos
Ascomicetos/química , Quitina/química , Proteínas Fúngicas/química , Hifas/química , Multimerização Proteica , Ascomicetos/genética , Ascomicetos/metabolismo , Quitina/genética , Quitina/metabolismo , Cladosporium/química , Cladosporium/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Hifas/genética , Hifas/metabolismo , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Estrutura Quaternária de Proteína , Triticum/genética , Triticum/metabolismo , Triticum/microbiologia
17.
Int J Food Microbiol ; 331: 108712, 2020 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-32563775

RESUMO

The bread-making quality of wheat depends on the viscoelastic properties of the dough in which gluten proteins play an important role. The quality of gluten proteins is influenced by the genetics of the different wheat varieties and environmental factors. Occasionally, a near complete loss of gluten strength, measured as the maximum resistance towards stretching (Rmax), is observed in grain lots of Norwegian wheat. It is hypothesized that the loss of gluten quality is caused by degradation of gluten proteins by fungal proteases. To identify fungi associated with loss of gluten strength, samples from a selection of wheat grain lots with weak gluten (n = 10, Rmax < 0.3 N) and strong gluten (n = 10, Rmax ≥ 0.6 N) was analyzed for the abundance of fungal operational taxonomic units (OTUs) using DNA metabarcoding of the nuclear ribosomal Internal Transcribed Spacer (ITS) region ITS1. The DNA quantities for a selection of fungal pathogens of wheat, and the total amount of fungal DNA, were analyzed by quantitative PCR (qPCR). The mean level of total fungal DNA was higher in grain samples with weak gluten compared to grain samples with strong gluten. Heightened quantities of DNA from fungi within the Fusarium Head Blight (FHB) complex, i.e. Fusarium avenaceum, Fusarium graminearum, Microdochium majus, and Microdochium nivale, were observed in grain samples with weak gluten compared to those with strong gluten. Microdochium majus was the dominant fungus in the samples with weak gluten. Stepwise regression modeling based on different wheat quality parameters, qPCR data, and the 35 most common OTUs revealed a significant negative association between gluten strength and three OTUs, of which the OTU identified as M. majus was the most abundant. The same analysis also revealed a significant negative relationship between gluten strength and F. avenaceum detected by qPCR, although the DNA levels of this fungus were low compared to those of M. majus. In vitro growth rate studies of a selection of FHB species showed that all the tested isolates were able to grow with gluten as a sole nitrogen source. In addition, proteins secreted by these fungi in liquid cultures were able to hydrolyze gluten substrate proteins in zymograms, confirming their capacity to secrete gluten-degrading proteases. The identification of fungi with potential to influence gluten quality can enable the development of strategies to minimize future problems with gluten strength in food-grade wheat.


Assuntos
Microbiologia de Alimentos , Fungos/classificação , Glutens/química , Triticum/química , Triticum/microbiologia , DNA Fúngico/genética , Grão Comestível/microbiologia , Fungos/genética , Fungos/isolamento & purificação , Fungos/metabolismo , Glutens/metabolismo , Doenças das Plantas/microbiologia , Reação em Cadeia da Polimerase em Tempo Real , Triticum/metabolismo
18.
Int J Food Microbiol ; 327: 108652, 2020 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-32442778

RESUMO

Food-grade waste and side streams should be strictly kept in food use in order to achieve sustainable food systems. At present, the baking industry creates food-grade waste as excess and deformed products that are mainly utilized for non-food uses, such as bioethanol production. The purpose of this study was therefore to explore the potential of waste wheat bread recycling for fresh wheat bread production. Waste bread recycling was assessed without further processing or after tailored fermentation with lactic acid bacteria producing either dextran or ß-glucan exopolysaccharides. When non-treated waste bread slurry was added to new bread dough, bread quality (specific volume and softness) decreased with increasing content of waste bread addition. In situ EPS-production (dextran and microbial ß-glucan) significantly increased waste bread slurry viscosity and yielded residual fructose or glucose that could effectively replace the sugar added for yeast leavening. Furthermore, fermentation acidified waste bread matrix, thus improving the hygienic safety of the process. Bread containing dextran synthesized in situ by Weissella confusa A16 showed good technological quality. The produced dextran compensated the adverse effect of recycled bread on new bread quality attributes by 12% increase in bread specific volume and 37% decrease in crumb hardness. In this study, a positive technological outcome of the bread containing microbial ß-glucan was not detected. The waste bread fermented by W. confusa A16 containing dextran appears to enable safe bread recycling with low acidity and minimal quality loss.


Assuntos
Pão/microbiologia , Pão/normas , Fermentação , Indústria Alimentícia , Resíduos Industriais , Reciclagem/métodos , Triticum/microbiologia , Dextranos/metabolismo , Ácido Láctico/metabolismo , Weissella/metabolismo , Leveduras/metabolismo , beta-Glucanas/metabolismo
19.
PLoS One ; 15(5): e0233878, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32470094

RESUMO

The present study aimed to investigate the effects of arbuscular mycorrhizal (AM) fungal communities originating from organic and conventional agriculture on wheat growth and yield. Six different spring wheat cultivars released in different years in north and central European countries were considered. We hypothesised that AM fungal inoculum collected from organic agricultural fields would elicit a greater positive growth response than inoculum collected from conventional agricultural fields; and that older cultivars, which were developed under conditions of low fertilizer input, would exhibit overall greater growth responses to the presence of AM fungi, compared with more recent cultivars, and that AM fungal inoculum from conventional fields might have the most beneficial effect on the growth and yield of recent cultivars. The results showed that the overall effects on the growth and yield of spring wheat grown with organic and conventional AM fungal inocula did not differ greatly. However, the inoculation growth response, showing the difference of the effects of organic and conventional inocula, varied between particular wheat cultivars. Inoculation growth response of the cultivar Pikker (released in 1959) was the most positive, while that of the cultivar Arabella (released in 2012) was the most negative. The use of AM fungal inoculum from organic fields resulted in slightly taller plant individuals. Pikker showed relatively higher yield and stronger growth when the organic AM fungal inoculum was used. Arabella exhibited relatively lower yield and weaker growth when the organic inoculum was used. Whether the positive response of Pikker to Estonian organic inoculation reflects adaptation to the locally occurring AM fungal community needs to be established by further studies of the communities of AM fungi colonizing wheat roots.


Assuntos
Fazendas , Micorrizas/fisiologia , Agricultura Orgânica , Triticum/crescimento & desenvolvimento , Triticum/microbiologia , Raízes de Plantas/fisiologia , Brotos de Planta/fisiologia , Solo , Microbiologia do Solo , Triticum/anatomia & histologia
20.
Int J Food Microbiol ; 324: 108631, 2020 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-32388172

RESUMO

In the present work the microbial dynamics in wheat kernels were evaluated over time. The main aim of this research was to study the resistance of lactic acid bacteria (LAB) and yeasts associated to unprocessed cereals used for bread making during long term conservation. To this purpose four Triticum durum Desf. genotypes including two modern varieties (Claudio and Simeto) and two Sicilian wheat landraces (Russello and Timilia) were analysed by a combined culture-independent and -dependent microbiological approach after one, two or three years from cultivation and threshing. DNA based MiSeq Illumina technology was applied to reveal the entire bacterial composition of all semolina samples. The samples showed a different distribution of bacterial taxa per variety and time of storage. The groups mostly represented were Stenotrophomonas, Pseudomonas, Erwinia, Delftia and Sphingomonas genera, Enterobacteriaceae and Oxalobacteriaceae families, and Actinobacteria phylum. Among LAB, only Enterococcus genus was detected barely in a single sample (Simeto stored for one year) by the next generation sequencing, indicating that LAB remained unassigned or their abundances were below 0.1% or their DNAs were rendered inaccessible. Plate counts showed consistent differences in relation to genotypes and duration of storage, with the highest levels found for total mesophilic microorganisms detected up 6.8 Log CFU/g. Colonies of presumptive sourdough LAB were detected only in a few samples. Cocci constituted the major group of LAB in almost all samples. Following the enrichment procedure, almost all samples were characterised by the presence of acidifying microorganisms. All isolates collected before and after enrichment represented 28 different strains belonging to 10 species of Enterococcus, Lactobacillus and Pediococcus genera. The most resistant species during aging were Enterococcus faecium, Enterococcus durans, Lactobacillus brevis, Lactobacillus pentosus and Lactobacillus paracasei demonstrating that lactobacilli and enterococci are able to overcome the stressing conditions represented by cereal storage better than other LAB genera commonly found associated to cereals after harvest. Yeast community included mostly species with no interest in bread making.


Assuntos
Pão/microbiologia , Grão Comestível/microbiologia , Microbiota/genética , Triticum/microbiologia , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Microbiologia de Alimentos , Armazenamento de Alimentos , Genótipo , Fatores de Tempo , Triticum/genética , Leveduras/genética , Leveduras/isolamento & purificação
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