RESUMO
In Europe, two fastidious phloem-limited pathogens, 'Candidatus Phytoplasma solani' (16SrXII-A) and 'Candidatus Arsenophonus phytopathogenicus', are associated with rubbery taproot disease (RTD) and syndrome basses richesses (SBR) of sugar beet, respectively. Both diseases can significantly reduce yield, especially when accompanied by root rot fungi. This study investigates the presence, geographic distribution and genetic traits of fastidious pathogens and the accompanying fungus, Macrophomina phaseolina, found on sugar beet across four geographically separated plains spanning seven countries in Central Europe. The survey revealed variable incidences of symptoms linked to these fastidious pathogens in the Pannonian and Wallachian Plains, sporadic occurrence in the North European Plain, and no symptomatic sugar beet in the Bohemian Plain. Molecular analyses unveiled the occurrence of both 'Ca. P. solani' and 'Ca. A. phytopathogenicus' throughout Central Europe, with a predominance of the phytoplasma. These fastidious pathogens were detected in all six countries surveyed within the Pannonian and Wallachian Plains, with only a limited presence of various phytoplasmas was found in the North European Plain, while no fastidious pathogens were detected in Bohemia, aligning with observed symptoms. While 16S rDNA sequences of 'Ca. P. solani' remained highly conserved, multi-locus characterization of two more variable loci (tuf and stamp) unveiled distinct variability patterns across the plains. Notably, the surprising lack of variability of tuf and stamp loci within Central Europe, particularly the Pannonian Plain, contrasted their high variability in Eastern and Western Europe, corresponding to epidemic and sporadic occurrence, respectively. The current study provides valuable insights into the genetic dynamics of 'Ca. P. solani' in Central Europe, and novel findings of the presence of 'Ca. A. phytopathogenicus' in five countries (Slovakia, Czech Republic, Austria, Serbia, and Romania) and M. phaseolina in sugar beet in Slovakia. These findings emphasize the need for further investigation of vector-pathogen(s)-plant host interactions and ecological drivers of disease outbreaks.
Assuntos
Beta vulgaris , Floema , Phytoplasma , Doenças das Plantas , Beta vulgaris/microbiologia , Europa (Continente)/epidemiologia , Doenças das Plantas/microbiologia , Phytoplasma/genética , Phytoplasma/patogenicidade , Phytoplasma/isolamento & purificação , Floema/microbiologia , Filogenia , Ascomicetos/genética , Geografia , PrevalênciaRESUMO
High boron (B) stress degrades the soil environment and reduces plant productivity. Sugar beet has a high B demand and potential for remediation of B-toxic soils. However, the mechanism regarding the response of sugar beet plants and rhizosphere soil microbiome to high B stress is not clear. In the potted soil experiment, we set different soil effective B environments (0.5, 5, 10, 30, 50, and 100 mg kg-1) to study the growth status of sugar beets under different B concentrations, as well as the characteristics of soil enzyme activity and microbial community changes. The results showed that sugar beet growth was optimal at 5 mg kg-1 of B. Exceeding this concentration the tolerance index decreased. The injury threshold EC20 was reached at an available B concentration of 35.8 mg kg-1. Under the treatment of 100 mg kg-1, the B accumulation of sugar beet reached 0.22 mg plant-1, and the tolerance index was still higher than 60%, which had not yet reached the lethal concentration of sugar beet. The abundance of Acidobacteriota, Chloroflexi and Patescibacteria increased, which was beneficial to the resistance of sugar beet to high B stress. In summary, under high B stress sugar beet had strong tolerance, enhanced capacity for B uptake and enrichment, and changes in soil microbial community structure. This study provides a theoretical basis for clarifying the mechanism of sugar beet resistance to high B stress and soil remediation.
Assuntos
Beta vulgaris , Solo , Solo/química , Beta vulgaris/metabolismo , Beta vulgaris/microbiologia , Boro , Rizosfera , Verduras , Açúcares/metabolismoRESUMO
BACKGROUND: Dietary fibers with varying physicochemical properties have different fermentation characteristics, which may differently impact host health. The present study aimed to determine the fermentation characteristics including gas production kinetics, short-chain fatty acids (SCFAs) production and microbial composition of different fibrous ingredients using in vitro fermentation by fecal microbiota. RESULTS: Sugar beet pule (SBP), wheat bran (WB), dried corn distillers grains with solubles (DDGS), rice bran (RB) and alfalfa meal (AM) were selected to fermentation in vitro for 36 h. The results showed that SBP had the greatest gas production. SBP had the highest in vitro dry matter fermentability (IVDMF) and production of acetate, propionate and total SCFAs, followed by WB, which were all greater than DDGS, AM and RB. The alpha-diversity was higher in the DDGS, AM and RB groups than in the WB and SBP groups. Differences in microbial community composition were observed among groups. The relative abundance of Treponema was highest in WB group. RB group showed lower Prevotella abundance than other groups but had higher Succinivibrio abundance. Interestingly, the Lactobacillus reached the highest abundances in the DDGS group. Correlation analysis indicated that the relative abundance of Treponema and Prevotella was positively associated with the gas production, IVDMF and SCFAs, whereas norank_f_Muribaculaceae, Rikenellaceae_RC9_gut_group, Lysinibacillus and Succinivibrio were the opposite. CONCLUSION: Collectively, WB and SBP were fermented rapidly by fecal microbiota compared to DDGS, AM and RB. Different fiber sources have different fiber compositions and fermentation properties that affect the microbial compositins and SCFAs production. © 2024 Society of Chemical Industry.
Assuntos
Ração Animal , Bactérias , Fibras na Dieta , Ácidos Graxos Voláteis , Fezes , Fermentação , Microbioma Gastrointestinal , Fibras na Dieta/metabolismo , Fibras na Dieta/análise , Fezes/microbiologia , Animais , Bactérias/metabolismo , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Ácidos Graxos Voláteis/metabolismo , Suínos , Ração Animal/análise , Zea mays/química , Zea mays/metabolismo , Beta vulgaris/química , Beta vulgaris/metabolismo , Beta vulgaris/microbiologia , Medicago sativa/química , Medicago sativa/metabolismo , Medicago sativa/microbiologia , Oryza/metabolismo , Oryza/química , Oryza/microbiologiaRESUMO
Cercospora leaf spot (CLS) is the most destructive foliar disease in sugar beet (Beta vulgaris). It is caused by Cercospora beticola Sacc., a fungal pathogen that produces toxins and enzymes which affect membrane permeability and cause cell death during infection. In spite of its importance, little is known about the initial stages of leaf infection by C. beticola. Therefore, we investigated the progression of C. beticola on leaf tissues of susceptible and resistant sugar beet varieties at 12-h intervals during the first 5 days after inoculation using confocal microscopy. Inoculated leaf samples were collected and stored in DAB (3,3'-diaminobenzidine) solution until processed. Samples were stained with Alexa Fluor-488-WGA dye to visualize fungal structures. Fungal biomass accumulation, reactive oxygen species (ROS) production, and the area under the disease progress curve were evaluated and compared. ROS production was not detected on any variety before 36 h postinoculation (hpi). C. beticola biomass accumulation, percentage leaf cell death, and disease severity were all significantly greater in the susceptible variety compared with the resistant variety (P < 0.05). Conidia penetrated directly through stomata between 48 to 60 hpi and produced appressoria on stomatal guard cells at 60 to 72 hpi in susceptible and resistant varieties, respectively. Penetration of hyphae inside the parenchymatous tissues varied in accordance with time postinoculation and varietal genotypes. Overall, this study provides a detailed account to date of events leading to CLS disease development in two contrasting varieties.
Assuntos
Ascomicetos , Beta vulgaris , Cercospora , Ascomicetos/fisiologia , Beta vulgaris/microbiologia , Espécies Reativas de Oxigênio , Suscetibilidade a Doenças , AçúcaresRESUMO
To investigate functional plant growth-promoting rhizobacteria in sugar beet, seasonal shifts in bacterial community structures in the lateral roots of sugar beet were examined using amplicon sequencing ana-lyses of the 16S rRNA gene. Shannon and Simpson indexes significantly increased between June and July, but did not significantly differ between July and subsequent months (August and September). A weighted UniFrac principal coordinate ana-lysis grouped bacterial samples into four clusters along with PC1 (43.8%), corresponding to the four sampling months in the order of sampling dates. Taxonomic ana-lyses revealed that bacterial diversity in the lateral roots was exclusively dominated by three phyla (Actinobacteria, Bacteroidetes, and Proteobacteria) in all samples examined. At the lower taxonomic levels, the dominant taxa were roughly classified into three groups. Therefore, the relative abundances of seven dominant genera (Janthinobacterium, Kribbella, Pedobacter, Rhodanobacter, Sphingobium, Sphingopyxis, and Streptomyces) were the highest in June and gradually decreased as sugar beet grew. The relative abundances of eight taxa (Bradyrhizobiaceae, Caulobacteraceae, Chitinophagaceae, Novosphingobium, Phyllobacteriaceae, Pseudomonas, Rhizobiaceae, and Sphingomonas) were mainly high in July and/or August. The relative abundances of six taxa (unclassified Comamonadaceae, Cytophagaceae, unclassified Gammaproteobacteria, Haliangiaceae, unclassified Myxococcales, and Sinobacteraceae) were the highest in September. Among the dominant taxa, 12 genera (Amycolatopsis, Bradyrhizobium, Caulobacter, Devosia, Flavobacterium, Janthinobacterium, Kribbella, Kutzneria, Pedobacter, Rhizobium, Rhodanobacter, and Steroidobacter) were considered to be candidate groups of plant growth-promoting bacteria based on their previously reported beneficial traits as biopesticides and/or biofertilizers.
Assuntos
Beta vulgaris , Beta vulgaris/microbiologia , RNA Ribossômico 16S/genética , Japão , Estações do Ano , Bactérias/genética , AçúcaresRESUMO
Fungal infections trigger defense or signaling responses in plants, leading to various changes in plant metabolites. The changes in metabolites, for example chlorophyll or flavonoids, have long been detectable using time-consuming destructive analytical methods including high-performance liquid chromatography or photometric determination. Recent plant phenotyping studies have revealed that hyperspectral imaging (HSI) in the UV range can be used to link spectral changes with changes in plant metabolites. To compare established destructive analytical methods with new nondestructive hyperspectral measurements, the interaction between sugar beet leaves and the pathogens Cercospora beticola, which causes Cercospora leaf spot disease (CLS), and Uromyces betae, which causes sugar beet rust (BR), was investigated. With the help of destructive analyses, we showed that both diseases have different effects on chlorophylls, carotenoids, flavonoids, and several phenols. Nondestructive hyperspectral measurements in the UV range revealed different effects of CLS and BR on plant metabolites resulting in distinct reflectance patterns. Both diseases resulted in specific spectral changes that allowed differentiation between the two diseases. Machine learning algorithms enabled the differentiation between the symptom classes and recognition of the two sugar beet diseases. Feature importance analysis identified specific wavelengths important to the classification, highlighting the utility of the UV range. The study demonstrates that HSI in the UV range is a promising, nondestructive tool to investigate the influence of plant diseases on plant physiology and biochemistry.
Assuntos
Ascomicetos , Beta vulgaris , Ascomicetos/fisiologia , Beta vulgaris/microbiologia , Imageamento Hiperespectral , Doenças das Plantas/microbiologia , Verduras , AçúcaresRESUMO
Sugar beet crown and root rot caused by Rhizoctonia solani is a major yield constraint. Root rot is highly increased when R. solani and Leuconostoc mesenteroides co-infect roots. We hypothesized that the absence of plant cell-wall-degrading enzymes in L. mesenteroides and their supply by R. solani during close contact, causes increased damage. In planta root inoculation with or without cell-wall-degrading enzymes showed greater rot when L. mesenteroides was combined with cellulase (22 mm rot), polygalacturonase (47 mm), and pectin lyase (57 mm) versus these enzymes (0-26 mm), R. solani (20 mm), and L. mesenteroides (13 mm) individually. Carbohydrate analysis revealed increased simpler carbohydrates (namely glucose + galactose, and fructose) in the infected roots versus mock control, possibly due to the degradation of complex cell wall carbohydrates. Expression of R. solani cellulase, polygalacturonase, and pectin lyase genes during root infection corroborated well with the enzyme data. Global mRNAseq analysis identified candidate genes and highly co-expressed gene modules in all three organisms that might be critical in host plant defense and pathogenesis. Targeting R. solani cell-wall-degrading enzymes in the future could be an effective strategy to mitigate root damage during its interaction with L. mesenteroides.
Assuntos
Beta vulgaris/microbiologia , Leuconostoc mesenteroides/metabolismo , Rhizoctonia/enzimologia , Beta vulgaris/crescimento & desenvolvimento , Beta vulgaris/metabolismo , Parede Celular/metabolismo , Expressão Gênica/genética , Regulação da Expressão Gênica de Plantas/genética , Leuconostoc mesenteroides/patogenicidade , Defesa das Plantas contra Herbivoria/imunologia , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Imunidade Vegetal/genética , Raízes de Plantas/metabolismo , Raízes de Plantas/microbiologia , Rhizoctonia/patogenicidadeRESUMO
Cercospora leaf spot (CLS) is a globally important disease of sugar beet (Beta vulgaris) caused by the fungus Cercospora beticola. Long-distance movement of C. beticola has been indirectly evidenced in recent population genetic studies, suggesting potential dispersal via seed. Commercial sugar beet "seed" consists of the reproductive fruit (true seed surrounded by maternal pericarp tissue) coated in artificial pellet material. In this study, we confirmed the presence of viable C. beticola in sugar beet fruit for 10 of 37 tested seed lots. All isolates harbored the G143A mutation associated with quinone outside inhibitor resistance, and 32 of 38 isolates had reduced demethylation inhibitor sensitivity (EC50 > 1 µg/ml). Planting of commercial sugar beet seed demonstrated the ability of seedborne inoculum to initiate CLS in sugar beet. C. beticola DNA was detected in DNA isolated from xylem sap, suggesting the vascular system is used to systemically colonize the host. We established nuclear ribosomal internal transcribed spacer region amplicon sequencing using the MinION platform to detect fungi in sugar beet fruit. Fungal sequences from 19 different genera were identified from 11 different sugar beet seed lots, but Fusarium, Alternaria, and Cercospora were consistently the three most dominant taxa, comprising an average of 93% relative read abundance over 11 seed lots. We also present evidence that C. beticola resides in the pericarp of sugar beet fruit rather than the true seed. The presence of seedborne inoculum should be considered when implementing integrated disease management strategies for CLS of sugar beet in the future.
Assuntos
Beta vulgaris , Cercospora , Beta vulgaris/microbiologia , Frutas , Doenças das Plantas/microbiologia , Açúcares , VerdurasRESUMO
BACKGROUND: Bacterial community successions were surveyed during the processing stages of sugar production using high-throughput sequencing methods. Furthermore, the correlation between bacterial community and nitrate/nitrite content in beet sugar processing were investigated. RESULTS: In an analysis of the V3-V4 region of the 16S rDNA gene, 254 122 effective sequences were obtained from samples, which included sugar beet, cossettes, diffusion juice, second-phase diffusion juice, light juice and thick juice. The results showed that dominant genera included Pantoea, Pseudomonas, Leuconostoc and Burkholderia. Moreover, significant changes in bacterial communities were observed in samples. Regarding the relevant nitrogen metabolic potential, this study revealed communities with the ability for nitrate and nitrite metabolism. Furthermore, a shaking experiment involving diffusion juice and second-phase diffusion juice was performed, and results showed that the nitrate level declined 73% and 98% in 36 h, respectively. These results suggested that the bacterial communities contribute to nitrate and nitrite transformation. CONCLUSION: This study illustrated that the bacterial communities and their specific effects on the formation of nitrate and nitrite during beet sugar processing. The results presented the basic concept involving the nitrate- and nitrite-forming pathways directly related to the mechanism of bacterial community growth. This study could facilitate an understanding of the correlation between nitrite content and microorganisms to guide beet sugar manufacturers regarding the control of nitrite and nitrate content. © 2021 Society of Chemical Industry.
Assuntos
Bactérias/metabolismo , Beta vulgaris/química , Nitratos/análise , Nitritos/análise , Tubérculos/microbiologia , Açúcares/isolamento & purificação , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Beta vulgaris/microbiologia , Biotransformação , Manipulação de Alimentos , Sequenciamento de Nucleotídeos em Larga Escala , Nitratos/metabolismo , Nitritos/metabolismo , Tubérculos/química , Açúcares/químicaRESUMO
Cercospora leaf spot (CLS) caused by Cercospora beticola is a devastating foliar disease of sugar beet (Beta vulgaris), resulting in high yield losses worldwide. Mycoviruses are widespread fungi viruses and can be used as a potential biocontrol agent for fugal disease management. To determine the presence of mycoviruses in C. beticola, high-throughput sequencing analysis was used to determine the diversity of mycoviruses in 139 C. beticola isolates collected from major sugar beet production areas in China. The high-throughput sequencing reads were assembled and searched against the NCBI database using BLASTn and BLASTx. The results showed that the obtained 93 contigs were derived from eight novel mycoviruses, which were grouped into 3 distinct lineages, belonging to the families Hypoviridae, Narnaviridae and Botourmiaviridae, as well as some unclassified (-)ssRNA viruses in the order Bunyavirales and Mononegavirales. To the best of our knowledge, this is the first identification of highly diverse mycoviruses in C. beticola. The novel mycoviruses explored in this study will provide new viral materials to biocontrol Cercospora diseases. Future studies of these mycoviruses will aim to assess the roles of each mycovirus in biological function of C. beticola in the future.
Assuntos
Cercospora/virologia , Micovírus/classificação , Doenças das Plantas/microbiologia , Plantas/microbiologia , Sequência de Aminoácidos , Beta vulgaris/microbiologia , Biodiversidade , China , Micovírus/genética , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , Doenças das Plantas/virologiaRESUMO
In nature, filamentous fungi are exposed to diverse nutritional sources and changes in substrate availability. Conversely, in submerged cultures, mycelia are continuously exposed to the existing substrates, which are depleted over time. Submerged cultures are the preferred choice for experimental setups in laboratory and industry and are often used for understanding the physiology of fungi. However, to what extent the cultivation method affects fungal physiology, with respect to utilization of natural substrates, has not been addressed in detail. Here, we compared the transcriptomic responses of Aspergillus niger grown in submerged culture and solid culture, both containing sugar beet pulp (SBP) as a carbon source. The results showed that expression of CAZy (Carbohydrate Active enZyme)-encoding and sugar catabolic genes in liquid SBP was time dependent. Moreover, additional components of SBP delayed the A. niger response to the degradation of pectin present in SBP. In addition, we demonstrated that liquid cultures induced wider transcriptome variability than solid cultures. Although there was a correlation regarding sugar metabolic gene expression patterns between liquid and solid cultures, it decreased in the case of CAZyme-encoding genes. In conclusion, the transcriptomic response of A. niger to SBP is influenced by the culturing method, limiting the value of liquid cultures for understanding the behavior of fungi in natural habitats. IMPORTANCE Understanding the interaction between filamentous fungi and their natural and biotechnological environments has been of great interest for the scientific community. Submerged cultures are preferred over solid cultures at a laboratory scale to study the natural response of fungi to different stimuli found in nature (e.g., carbon/nitrogen sources, pH). However, whether and to what extent submerged cultures introduce variation in the physiology of fungi during growth on plant biomass have not been studied in detail. In this study, we compared the transcriptomic responses of Aspergillus niger to growth on liquid and solid cultures containing sugar beet pulp (a by-product of the sugar industry) as a carbon source. We demonstrate that the transcriptomic response of A. niger was highly affected by the culture condition, since the transcriptomic response obtained in a liquid environment could not fully explain the behavior of the fungus in a solid environment. This could partially explain the differences often observed between the phenotypes on plates compared to liquid cultures.
Assuntos
Aspergillus niger/crescimento & desenvolvimento , Aspergillus niger/genética , Beta vulgaris/microbiologia , Proteínas Fúngicas/genética , Aspergillus niger/metabolismo , Beta vulgaris/metabolismo , Meios de Cultura/química , Meios de Cultura/metabolismo , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Pectinas/metabolismo , TranscriptomaRESUMO
Sulfur is an essential plant macronutrient, and its adequate supply allows an efficient root storage and sugar extractability in sugar beets (Beta vulgaris L.). In this study, we investigated the effect of changes in sulfur availability on the endophytic community structure of sugar beets. Plants were hydroponically grown in a complete nutrient solution (S-supplied), a nutrient solution without MgSO4 (S-deprived), and a nutrient solution without MgSO4 for six days and resupplied with 100 µM MgSO4 for 48 h (S-resupplied). The sulfur status was monitored by inductively coupled plasma ICP-OES, and combustion analysis together with the evaluation of microRNA395 as a biomarker for sulfate status. Metabarcoding of the bacterial 16S rRNA gene was carried out in order to determine leaf endophytic community structure. The Shannon diversity index significantly differed (p < 0.05) between sulfate-supplied and sulfate-deprived seedlings. Validation by Real-Time PCR showed a significant increase (p < 0.05) of Burkholderia spp. in sulfate-deprived plants as compared to sulfate-supplied ones. The study sheds new light on the effects of nutrient deficiency on the microbiome of sugar beet plants.
Assuntos
Beta vulgaris/microbiologia , Endófitos , Microbiota , Enxofre , MetagenomaRESUMO
Clone libraries of bacterial 16S rRNA genes (a total of 1,980 clones) were constructed from the leaf blades, petioles, taproots, and lateral roots of sugar beet (Beta vulgaris L.) grown under different fertilization conditions. A principal coordinate analysis revealed that the structures of bacterial communities in above- and underground tissues were largely separated by PC1 (44.5%). The bacterial communities of above-ground tissues (leaf blades and petioles) were more tightly clustered regardless of differences in the tissue types and fertilization conditions than those of below-ground tissues (taproots and lateral roots). The bacterial communities of below-ground tissues were largely separated by PC2 (26.0%). To survey plant growth-promoting bacteria (PGPBs), isolate collections (a total of 665 isolates) were constructed from the lateral roots. As candidate PGPBs, 44 isolates were selected via clustering analyses with the combined 16S rRNA gene sequence data of clone libraries and isolate collections. The results of inoculation tests using sugar beet seedlings showed that eight isolates exhibited growth-promoting effects on the seedlings. Among them, seven isolates belonging to seven genera (Asticcacaulis, Mesorhizobium, Nocardioides, Sphingobium, Sphingomonas, Sphingopyxis, and Polaromonas) were newly identified as PGPBs for sugar beet at the genus level, and two isolates belonging to two genera (Asticcacaulis and Polaromonas) were revealed to exert growth-promoting effects on the plant at the genus level for the first time. These results suggest that a community analysis-based selection strategy will facilitate the isolation of novel PGPBs and extend the potential for the development of novel biofertilizers.
Assuntos
Bactérias/isolamento & purificação , Beta vulgaris/crescimento & desenvolvimento , Microbiota , Bactérias/classificação , Bactérias/genética , Beta vulgaris/microbiologia , DNA Bacteriano/genética , Folhas de Planta/microbiologia , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/microbiologia , RNA Ribossômico 16S/genética , Plântula/crescimento & desenvolvimento , Plântula/microbiologia , Microbiologia do SoloRESUMO
The increase of human population and associated increasing demand for agricultural products lead to soil over-exploitation. Biofertilizers based on lyophilized plant material containing living plant growth-promoting microorganisms (PGPM) could be an alternative to conventional fertilizers that fits into sustainable agricultural technologies ideas. We aimed to: (1) assess the diversity of endophytic bacteria in sugar and sea beet roots and (2) determine the influence of osmoprotectants (trehalose and ectoine) addition during lyophilization on bacterial density, viability and salt tolerance. Microbiome diversity was assessed based on 16S rRNA amplicons sequencing, bacterial density and salt tolerance was evaluated in cultures, while bacterial viability was calculated by using fluorescence microscopy and flow cytometry. Here we show that plant genotype shapes its endophytic microbiome diversity and determines rhizosphere soil properties. Sea beet endophytic microbiome, consisting of genera characteristic for extreme environments, is more diverse and salt resistant than its crop relative. Supplementing osmoprotectants during root tissue lyophilization exerts a positive effect on bacterial community salt stress tolerance, viability and density. Trehalose improves the above-mentioned parameters more effectively than ectoine, moreover its use is economically advantageous, thus it may be used to formulate improved biofertilizers.
Assuntos
Beta vulgaris/crescimento & desenvolvimento , Produção Agrícola/métodos , Endófitos/fisiologia , Microbiota/fisiologia , Microbiologia do Solo , Beta vulgaris/microbiologia , DNA Bacteriano/isolamento & purificação , Endófitos/isolamento & purificação , Liofilização , Humanos , RNA Ribossômico 16S/genética , Rizosfera , Tolerância ao Sal , Desenvolvimento SustentávelRESUMO
The fungal species Rhizoctonia solani belongs to the Basidiomycota division and is a ubiquitous soil-borne pathogen. It is the main agent of the damping-off disease in seedlings and causes the root and crown rot disease in sugar beets. Plant pathogens deploy small secreted proteins, called effectors, to manipulate plant immunity in order to infect the host. Here, a gene (RsCRP1) encoded a putative effector cysteine-rich protein was cloned, expressed in Cercospora beticola and used for virulence assays. The RsCRP1 gene was highly induced upon the early-infection stage of sugar beet seedlings and disease was promoted. Confocal microscopy demonstrated localization to the chloroplasts and mitochondria upon transient expression of RsCRP1 in leaves of Nicotiana benthamiana. Further, this effector was unable to induce necrosis or to suppress hypersensitive response induced by the Avr4/Cf4 complex in N. benthamiana. Overall, these data indicate that RsCRP1 is a novel effector targeting distinct plant cell organelles in order to facilitate a successful infection at the early stages of the disease development.
Assuntos
Beta vulgaris/crescimento & desenvolvimento , Cloroplastos/metabolismo , Mitocôndrias/metabolismo , Doenças das Plantas/microbiologia , Rhizoctonia/patogenicidade , Plântula/crescimento & desenvolvimento , Fatores de Virulência/metabolismo , Beta vulgaris/metabolismo , Beta vulgaris/microbiologia , Cloroplastos/microbiologia , Mitocôndrias/microbiologia , Doenças das Plantas/genética , Imunidade Vegetal , Folhas de Planta/metabolismo , Folhas de Planta/microbiologia , Plântula/metabolismo , Plântula/microbiologia , Nicotiana/metabolismo , Nicotiana/microbiologiaRESUMO
Cercospora beticola is a hemibiotrophic fungus that causes cercospora leaf spot disease of sugar beet (Beta vulgaris). After an initial symptomless biotrophic phase of colonization, necrotic lesions appear on host leaves as the fungus switches to a necrotrophic lifestyle. The phytotoxic secondary metabolite cercosporin has been shown to facilitate fungal virulence for several Cercospora spp. However, because cercosporin production and subsequent cercosporin-initiated formation of reactive oxygen species is light-dependent, cell death evocation by this toxin is only fully ensured during a period of light. Here, we report the discovery of the effector protein CbNip1 secreted by C. beticola that causes enhanced necrosis in the absence of light and, therefore, may complement light-dependent necrosis formation by cercosporin. Infiltration of CbNip1 protein into sugar beet leaves revealed that darkness is essential for full CbNip1-triggered necrosis, as light exposure delayed CbNip1-triggered host cell death. Gene expression analysis during host infection shows that CbNip1 expression is correlated with symptom development in planta. Targeted gene replacement of CbNip1 leads to a significant reduction in virulence, indicating the importance of CbNip1 during colonization. Analysis of 89 C. beticola genomes revealed that CbNip1 resides in a region that recently underwent a selective sweep, suggesting selection pressure exists to maintain a beneficial variant of the gene. Taken together, CbNip1 is a crucial effector during the C. beticola-sugar beet disease process.
Assuntos
Beta vulgaris/microbiologia , Cercospora/genética , Proteínas Fúngicas/metabolismo , Genoma Fúngico/genética , Perileno/análogos & derivados , Doenças das Plantas/microbiologia , Cercospora/crescimento & desenvolvimento , Cercospora/patogenicidade , Proteínas Fúngicas/genética , Interações Hospedeiro-Patógeno , Necrose , Perileno/metabolismo , Fenótipo , Filogenia , Folhas de Planta/microbiologia , Virulência , Fatores de VirulênciaRESUMO
BACKGROUND: Exhausted sugar beet pulp pellets (ESBPP) were used as raw material for lactic acid (LA) fermentation. The enzymatic hydrolysis of ESBPP was performed with the solid obtained after the fungal solid-state fermentation of ESBPP as a source of hydrolytic enzymes. Subsequently, a medium rich in glucose and arabinose was obtained, which was used to produce LA by fermentation. For LA production, two Lactobacillus strains were assayed and the effects of the supplementation of the hydrolysate with a nitrogen source and the mode of pH regulation of the fermentation were investigated. Moreover, a kinetic model for LA fermentation by Lactobacillus plantarum of ESBPP hydrolysates was developed. RESULTS: L. plantarum produced a LA concentration 34% higher than that produced by L. casei. The highest LA concentration (30 g L-1 ) was obtained with L. plantarum when the hydrolysate was supplemented with 5 g L-1 yeast extract and the pH was controlled with CaCO3 . The concentration of acetic acid differed depending on the concentration of CaCO3 added, producing its maximum value with 27 g L-1 CaCO3 . The proposed kinetic model was able to predict the evolution of substrates and products depending on the variation of the pH in the hydrolysate, according to the amount of CaCO3 added. CONCLUSIONS: ESBPP can be revalorised to produce LA. A pure LA stream or a mixture of LA and acetic acid, depending on the pH control method of the fermentation, can be produced. Thus, this control is of great interest depending on the destination of the effluent. © 2020 Society of Chemical Industry.
Assuntos
Beta vulgaris/microbiologia , Meios de Cultura/metabolismo , Fungos/metabolismo , Ácido Láctico/biossíntese , Lactobacillus/metabolismo , Ácido Acético/metabolismo , Beta vulgaris/química , Beta vulgaris/metabolismo , Meios de Cultura/análise , Fermentação , Cinética , Resíduos/análiseRESUMO
Sugar beets are attacked by several pathogens that cause root damages. Rhizoctonia (Greek for "root killer") is one of them. Rhizoctonia root rot has become an increasing problem for sugar beet production and to decrease yield losses agronomical measures are adopted. Here, two partially resistant and two susceptible sugar beet genotypes were used for transcriptome analysis to discover new defense genes to this fungal disease, information to be implemented in molecular resistance breeding. Among 217 transcripts with increased expression at 2 days post-infection (dpi), three resistance-like genes were found. These genes were not significantly elevated at 5 dpi, a time point when increased expression of three Bet v I/Major latex protein (MLP) homologous genes BvMLP1, BvMLP2 and BvML3 was observed in the partially resistant genotypes. Quantitative RT-PCR analysis on diseased sugar beet seedlings validated the activity of BvMLP1 and BvMLP3 observed in the transcriptome during challenge by R. solani. The three BvMLP genes were cloned and overexpressed in Arabidopsis thaliana to further dissect their individual contribution. Transgenic plants were also compared to T-DNA mutants of orthologous MLP genes. Plants overexpressing BvMLP1 and BvMLP3 showed significantly less infection whereas additive effects were seen on Atmlp1/Atmlp3 double mutants. The data suggest that BvMLP1 and BvMLP3 may contribute to the reduction of the Rhizoctonia root rot disease in sugar beet. Impact on the defense reaction from other differential expressed genes observed in the study is discussed.
Assuntos
Beta vulgaris/genética , Regulação da Expressão Gênica de Plantas/imunologia , Doenças das Plantas/genética , Proteínas de Plantas/genética , Rhizoctonia/patogenicidade , Transcriptoma/imunologia , Arabidopsis/genética , Arabidopsis/metabolismo , Beta vulgaris/imunologia , Beta vulgaris/microbiologia , Clonagem Molecular , Expressão Gênica , Redes Reguladoras de Genes , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Imunidade Vegetal/genética , Proteínas de Plantas/imunologia , Plantas Geneticamente Modificadas , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Rhizoctonia/crescimento & desenvolvimento , Plântula/genética , Plântula/imunologia , Plântula/microbiologiaRESUMO
BACKGROUND: Root and stem rot caused by Rhizoctonia solani is a serious fungal disease of sugar beet and dry bean production in Nebraska. Rhizoctonia root rot and crown rot in sugar beet and dry bean have reduced the yield significantly and has also created problems in storage. The objective of this study was to analyze morpho-genetic diversity of 38 Rhizoctonia solani isolates from sugar beet and dry bean fields in western Nebraska collected over 10 years. Morphological features and ISSR-based DNA markers were used to study the morphogenetic diversity. RESULTS: Fungal colonies were morphologically diverse in shapes, aerial hyphae formation, colony, and sclerotia color. Marker analysis using 19 polymorphic ISSR markers showed polymorphic bands ranged from 15 to 28 with molecular weight of 100 bp to 3 kb. Polymorphic loci ranged from 43.26-92.88%. Nei genetic distance within the population ranged from 0.03-0.09 and Shannon diversity index varied from 0.24-0.28. AMOVA analysis based on ΦPT values showed 87% variation within and 13% among the population with statistical significance (p < 0.05). Majority of the isolates from sugar beet showed nearby association within the population. A significant number of isolates showed similarity with isolates of both the crops suggesting their broad pathogenicity. Isolates were grouped into three different clusters in UPGMA based cluster analysis using marker information. Interestingly, there was no geographical correlation among the isolates. Principal component analysis showed randomized distribution of isolates from the same geographical origin. Identities of the isolates were confirmed by both ITS-rDNA sequences and pathogenicity tests. CONCLUSION: Identification and categorization of the pathogen will be helpful in designing integrated disease management guidelines for sugar beet and dry beans of mid western America.
Assuntos
Beta vulgaris/microbiologia , Phaseolus/microbiologia , Doenças das Plantas/microbiologia , Rhizoctonia/genética , Análise por Conglomerados , DNA Fúngico/genética , Marcadores Genéticos , Variação Genética , Estudos Longitudinais , Repetições de Microssatélites/genética , Nebraska , Raízes de Plantas/microbiologia , Rhizoctonia/classificação , Rhizoctonia/citologia , Rhizoctonia/isolamento & purificaçãoRESUMO
Rhizoctonia solani (Rs) is a soil-borne pathogen with a broad host range. This pathogen incites a wide range of disease symptoms. Knowledge regarding its infection process is fragmented, a typical feature for basidiomycetes. In this study, we aimed at identifying potential fungal effectors and their function. From a group of 11 predicted single gene effectors, a rare lipoprotein A (RsRlpA), from a strain attacking sugar beet was analyzed. The RsRlpA gene was highly induced upon early-stage infection of sugar beet seedlings, and heterologous expression in Cercospora beticola demonstrated involvement in virulence. It was also able to suppress the hypersensitive response (HR) induced by the Avr4/Cf4 complex in transgenic Nicotiana benthamiana plants and functioned as an active protease inhibitor able to suppress Reactive Oxygen Species (ROS) burst. This effector contains a double-psi beta-barrel (DPBB) fold domain, and a conserved serine at position 120 in the DPBB fold domain was found to be crucial for HR suppression. Overall, R. solani seems to be capable of inducing an initial biotrophic stage upon infection, suppressing basal immune responses, followed by a switch to necrotrophic growth. However, regulatory mechanisms between the different lifestyles are still unknown.