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Microorganisms Dec 2022, the causative agent of fire blight, leads to important economic losses of apple and pear crops worldwide. This study aimed to investigate the potential of the resident...
, the causative agent of fire blight, leads to important economic losses of apple and pear crops worldwide. This study aimed to investigate the potential of the resident microbiota of the apple blossom in combatting plant disease-causing organisms, with a focus on controlling fire blight. We obtained 538 isolates from sites around Canton Zurich, which we tested for activity against and We also evaluated the isolates' activity against oomycete and fungal pathogens. Nine isolates showed activity against , and eight of these against . Furthermore, 117 showed antifungal, and 161 anti-oomycete, activity. We assigned genera and in some cases species to 238 of the isolates by sequencing their 16S RNA-encoding gene. Five strains showed activity against all pathogens and were tested in a detached apple model for anti- activity. Of these five strains, two were able to antagonize , namely #124 and #378. We sequenced the #378 genome and analyzed it for secondary metabolite clusters using antiSMASH, revealing the presence of a putative bacteriocin cluster. We also showed that #124 exhibits strong activity against three different fungi and two oomycetes in vitro, suggesting a broader capacity for biocontrol. Our results showcase the protective potential of the natural apple blossom microbiota. We isolated two candidate biocontrol strains from apple blossoms, suggesting that they might persist at the most common entry point for the causative agent of fire blight. Furthermore, they are probably already part of the human diet, suggesting they might be safe for consumption, and thus are promising candidates for biocontrol applications.
PubMed: 36557734
DOI: 10.3390/microorganisms10122480 -
Journal of Applied Microbiology Jan 2007To investigate the aetiology of seed and boll rot of cotton grown in South Carolina (SC).
AIMS
To investigate the aetiology of seed and boll rot of cotton grown in South Carolina (SC).
METHODS AND RESULTS
Bacteria were isolated from diseased locules of cotton bolls collected in a field in SC, USA and tested for the ability to cause comparable disease symptoms in greenhouse grown cotton fruit. Spontaneously generated rifampicin-resistant (Rif(r)) mutants of the isolates were used in confirmatory pathogenicity tests. Resistance to the antibiotic was both stable and effective in differentiating between an inoculated Rif(r) strain, rifampicin-sensitive contaminants and/or endophytes. A series of inoculation methods was tested at various boll developmental stages and at different fruiting nodes on the plant. Field disease symptoms were reproduced by inoculating bolls at 2 weeks postanthesis with bacterial suspensions ranging from 10(3) to 10(6) CFU ml(-1). Pathogenic isolates were categorized as Pantoea agglomerans on the basis of phenotype testing, fatty acid profiling (similarity index = 0.94), and 16s ribosomal DNA sequence analysis (99% nucleotide identity).
CONCLUSIONS
Pantoea agglomerans isolates from field-collected immature, diseased cotton caused comparable infection symptoms in greenhouse produced cotton fruit.
SIGNIFICANCE AND IMPACT OF THE STUDY
In 1999, significant yield losses in SC cotton resulted from a previously unobserved seed and boll rot that has since been reported in other southeastern states. This study demonstrated a role of P. agglomerans in producing opportunistic bacterial seed and boll rot of cotton.
Topics: Anti-Bacterial Agents; Base Sequence; DNA, Bacterial; DNA, Ribosomal; Drug Resistance, Bacterial; Fatty Acids; Gossypium; Gram-Negative Bacterial Infections; Pantoea; Phenotype; Plant Diseases; Rifampin; South Carolina
PubMed: 17184328
DOI: 10.1111/j.1365-2672.2006.03055.x -
International Journal of Molecular... Jul 2021A novel siphovirus, vB_PagS_MED16 (MED16) was isolated in Lithuania using strain BSL for the phage propagation. The double-stranded DNA genome of MED16 (46,103 bp)...
A novel siphovirus, vB_PagS_MED16 (MED16) was isolated in Lithuania using strain BSL for the phage propagation. The double-stranded DNA genome of MED16 (46,103 bp) contains 73 predicted open reading frames (ORFs) encoding proteins, but no tRNA. Our comparative sequence analysis revealed that 26 of these ORFs code for unique proteins that have no reliable identity when compared to database entries. Based on phylogenetic analysis, MED16 represents a new genus with siphovirus morphology. In total, 35 MED16 ORFs were given a putative functional annotation, including those coding for the proteins responsible for virion morphogenesis, phage-host interactions, and DNA metabolism. In addition, a gene encoding a preQ DNA deoxyribosyltransferase (DpdA) is present in the genome of MED16 and the LC-MS/MS analysis indicates 2'-deoxy-7-amido-7-deazaguanosine (dADG)-modified phage DNA, which, to our knowledge, has never been experimentally validated in genomes of phages. Thus, the data presented in this study provide new information on -infecting viruses and offer novel insights into the diversity of DNA modifications in bacteriophages.
Topics: DNA, Viral; Genome, Viral; Guanosine; Open Reading Frames; Pantoea; Siphoviridae; Viral Proteins
PubMed: 34298953
DOI: 10.3390/ijms22147333 -
Applied and Environmental Microbiology Jan 2011A bacterium capable of producing a deep blue pigment was isolated from the environment and identified as Pantoea agglomerans. The pigment production characteristics of...
A bacterium capable of producing a deep blue pigment was isolated from the environment and identified as Pantoea agglomerans. The pigment production characteristics of the bacterium under various conditions were studied. The optimal agar plate ingredients for pigment production by the bacterium were first studied: the optimal ingredients were 5 g/liter glucose, 10 g/liter tryptic soy broth, and 40 g/liter glycerol at pH 6.4. Bacterial cells grew on the agar plate during the incubation, while the pigment spread into the agar plate, meaning that it is water soluble. Pigment production was affected by the initial cell density. Namely, at higher initial cell densities ranging from 10(6.3) to 10(8.2) CFU/cm(2) on the agar plate, faster pigment production was observed, but no blue pigment was produced at a very high initial density of 10(9.1) CFU/cm(2). Thus, the cell population of 10(8.2) CFU/cm(2) was used for subsequent study. Although the bacterium was capable of growing at temperatures above and below 10°C, it could produce the pigment only at temperatures of ≥10°C. Moreover, the pigment production was faster at higher temperatures in the range of 10 to 20°C. Pigment production at various temperature patterns was well described by a new logistic model. These results suggested that the bacterium could be used in the development of a microbial temperature indicator for the low-temperature-storage management of foods and clinical materials. To our knowledge, there is no other P. agglomerans strain capable of producing a blue pigment and the pigment is a new one of microbial origin.
Topics: Culture Media; Environmental Microbiology; Pantoea; Pigments, Biological; Temperature
PubMed: 20971865
DOI: 10.1128/AEM.00264-10 -
Journal of Applied Microbiology Jul 2014This study aimed to evaluate different packaging strategies to extend the shelf life of a freeze-dried formulation of the biocontrol agent Pantoea agglomerans strain...
AIMS
This study aimed to evaluate different packaging strategies to extend the shelf life of a freeze-dried formulation of the biocontrol agent Pantoea agglomerans strain CPA-2.
METHODS AND RESULTS
Different materials and atmosphere packaging conditions (vacuum and air) were analysed on formulated P. agglomerans cells stored at 25, 5 and -20°C. Results showed the viability of CPA-2 cells stored at 5 or -20°C was significantly higher than when stored at 25°C. The highest viabilities were observed with the plastic material designated as Bottle 1, in nonvacuum packaging in all storage temperatures: 50% after 3 months at 25°C, 100% after 8 months at 5°C and 100 and 74% after 12 and 18 months, respectively, at -20°C; the final concentration was 10(12) CFU g(-1), a good concentration for a commercial product. The efficacy to control blue and green mould on apples and oranges, respectively, of these packed and stored cells was similar to fresh CPA-2 cells.
CONCLUSIONS
This work showed a suitable packaging strategy for a freeze-dried formulation of the CPA-2, providing a good shelf life and efficacy against the major postharvest diseases of apples and citrus based on a plastic bottle stored at cold or frozen storage conditions.
SIGNIFICANCE AND IMPACT OF THE STUDY
The last phase of the commercial development process for biocontrol agents is presented in this work. A bacterium-based product that ensures the efficacy, stability and easy application of the antagonist to control postharvest fungal diseases on fruit was successfully obtained.
Topics: Biological Control Agents; Citrus sinensis; Food Packaging; Food Storage; Freeze Drying; Fruit; Humans; Malus; Pantoea; Refrigeration
PubMed: 24698363
DOI: 10.1111/jam.12511 -
Plant Disease Dec 2006In April 2006, sweet onions (Allium cepa) that were grown in Wayne County, GA displayed symptoms typical of either center rot caused by Pantoea ananatis or a foliar...
In April 2006, sweet onions (Allium cepa) that were grown in Wayne County, GA displayed symptoms typical of either center rot caused by Pantoea ananatis or a foliar blight caused by Iris yellow spot virus (IYSV). After samples tested negative for IYSV by enzyme-linked immunosorbent assay and polymerase chain reaction, isolations were made from basal areas of leaves of infected plants where healthy and diseased tissues converged. All samples yielded yellow colonies on trypticase soy broth agar (TSBA) that were nonfluorescent when transferred to King's medium B. Four strains were characterized and tentatively identified as a Pantoea sp. by yellow pigmentation of colonies, oxidative and fermentative use of glucose, and lack of oxidase. However, the inability to produce indole from tryptophan, negative ice-nucleation activity, ability to reduce nitrate to nitrite, and the presence of phenylalanine deaminase were characteristics more typical of P. agglomerans than P. ananatis. Furthermore, all test strains utilized cellobiose, raffinose, lactose, gelatin, melibiose, and malonate. The identity of the bacterium was confirmed as P. agglomerans by BIOLOG (Hayward, CA). In addition, the 16S gene was amplified using universal primers (forward 5'-AGTTTGATCCTGGCTCAG-3' and reverse 5'-TACCTTGTTACGACTTCGTCCCA-3' (1) and sequenced. A BLAST search of the sequence against the NIH GenBank nucleotide library also confirmed the identity of the onion pathogen as P. agglomerans (97% identity) by having 8 of the top 10 bacteria providing significant alignments identified as P. agglomerans. The remaining two matches were uncultured bacteria from environmental samples. To confirm pathogenicity, two onion plants for each of the four test strains were inoculated with a turbid, aqueous bacterial suspension (~1 × 10 CFU ml) or sterile water in the lab (n = 8) and the field (n = 8). In addition, two plants each were inoculated with P. ananatis as a positive control and with a water blank and a nonpathogenic strain of P. agglomerans from peach (Png 86-2) as negative controls. All test strains of P. agglomerans produced severe blighting and withering of onion leaves in 4 days, while the water control and Png 86-2 were negative. Results were the same for both lab and field trials. Bacteria recovered from the plants infected with the test strains demonstrated the same characteristics of P. agglomerans as described above. Although P. agglomerans was originally reported as a pathogen of onion in South Africa (2), to the best of our knowledge, this is the first report of P. agglomerans causing a disease of onions in the United States. The long-term impact on the onion industry at this time is unknown. However, considering the close relationship of this organism with P. ananatis and the similarity of disease symptoms with those caused by center rot, there is potential that this bacterium could become established in the onion-growing area of Georgia and become part of a center rot 'complex'. References: (1) T. De Baere et al. J. Clin. Microbiol. 42:4393, 2004. (2) M. J. Hattingh and D. F. Walters. Plant Dis. 65:615, 1981.
PubMed: 30780978
DOI: 10.1094/PD-90-1551A -
Plant Disease Oct 2007Zea mays and Sorghum bicolor are important crops for animal and human nutrition worldwide. In the Central Highland Valley of Mexico, both crops are extremely important,...
Zea mays and Sorghum bicolor are important crops for animal and human nutrition worldwide. In the Central Highland Valley of Mexico, both crops are extremely important, and research is aimed toward increasing yield, disease resistance, and crop adaptation from 1,900- to 2,700-m elevation. In a 3-year field breeding experiment (2004 to 2006), leaf blight and vascular wilt symptoms were frequently observed in contiguous plots of maize and sorghum crops in Montecillo, Mexico and maize plots in Tecamac, Mexico. To identify and characterize the causal agent of these symptoms, isolations were conducted on leaves from areas where healthy and diseased tissues converged. Leaf sections of 1 cm2 from both crops were disinfested, placed on casamino acid-peptone-glucose (CPG) medium, and incubated at 28°C. After 48 h, only yellow colonies were observed and 12 isolates were selected for further characterization. Physiological and biochemical tests indicated that the isolates were nonfluorescent on King's B medium, and API 50 CHE (bioMérieux, Marcy l'Etoile, France) revealed that they were negative for gelatin hydrolysis, indole production, acid production from raffinose and positive for utilization of glycerol, D-glucose, mannitol, arbutine, esculine, salicine, cellobiose, maltose, melibiose, D-fucose, and D-arabitol; all characteristics of Pantoea agglomerans. Further identification of these isolates was accomplished by DNA analysis. For DNA analysis, 1.4-kbp fragments of the 16S rRNA gene were amplified with primer set 8F/1492R (3) and sequenced with U514F/800R universal primers (2). Five sequences were obtained and deposited in GenBank (Accession Nos. EF050806 to EF050810). A phylogenetic tree was constructed using the UPGMA method (mega version 3.1). Results of the phylogenetic analysis grouped the species P. ananatis, P. stewartti, and P. agglomerans into three clusters. The five unknown sequences were grouped into the P. agglomerans cluster. There was a 98 to 99% similarity of the five 16S rRNA gene sequences with P. agglomerans strain type ATCC 27155. Pathogenicity of the 12 isolates was confirmed by injecting 10 CFU mL of inoculum into stems of 3-week-old maize cv. Triunfo and sorghum cold tolerant hybrid (A1×B5)×R1 seedlings in the greenhouse at 28°C and 80% relative humidity. Also, seedlings were inoculated with water, nonpathogenic isolates of P. agglomerans from maize (GM13, and HLA1), and not inoculated as negative controls. Three replications were included for each isolate and control. All test strains developed water-soaked lesions on juvenile leaves at 8 days postinoculation and were followed by chlorotic to straw-colored leaf streaks and then leaf blight symptoms at 3 weeks postinoculation. All negative control seedlings did not develop symptoms. In addition, the 12 isolates were infiltrated at 10 CFU mL into tobacco leaves that displayed a hypersensitive response at 4 days, indicating the presence of the type III secretion system (1). Isolates were reisolated, and the 16S rRNA gene fragments were 100% similar to their original isolate sequences. P. agglomerans has been reported to affect other crops, including chinese taro in Brazil (2007), onion in the United States (2006) and South Africa (1981), and pearl millet in Zimbabwe (1997); however, to our knowledge, this is the first report of P. agglomerans associated with leaf blight and vascular wilt symptoms in maize and sorghum in the Central Highland Valley of Mexico. References: (1) J. Alfano and A. Collmer. Annu. Rev. Phytopathol 42:385, 2004. (2) Y. Anzai et al. Int. J. Syst. Evol. Microbiol. 50:1563, 2000. (3) M. Sasoh et al. Appl. Environ. Microbiol. 72:1825, 2006.
PubMed: 30780539
DOI: 10.1094/PDIS-91-10-1365A -
Frontiers in Plant Science 2015There is an increasing interest in studying interspecies bacterial interactions in diseases of animals and plants as it is believed that the great majority of bacteria... (Review)
Review
There is an increasing interest in studying interspecies bacterial interactions in diseases of animals and plants as it is believed that the great majority of bacteria found in nature live in complex communities. Plant pathologists have thus far mainly focused on studies involving single species or on their interactions with antagonistic competitors. A bacterial disease used as model to study multispecies interactions is the olive knot disease, caused by Pseudomonas savastanoi pv. savastanoi (Psv). Knots caused by Psv in branches and other aerial parts of the olive trees are an ideal niche not only for the pathogen but also for many other plant-associated bacterial species, mainly belonging to the genera Pantoea, Pectobacterium, Erwinia, and Curtobacterium. The non-pathogenic bacterial species Erwinia toletana, Pantoea agglomerans, and Erwinia oleae, which are frequently isolated inside the olive knots, cooperate with Psv in modulating the disease severity. Co-inoculations of these species with Psv result in bigger knots and better bacterial colonization when compared to single inoculations. Moreover, harmless bacteria co-localize with the pathogen inside the knots, indicating the formation of stable bacterial consortia that may facilitate the exchange of quorum sensing signals and metabolites. Here we discuss the possible role of bacterial communities in the establishment and development of olive knot disease, which we believe could be taking place in many other bacterial plant diseases.
PubMed: 26113855
DOI: 10.3389/fpls.2015.00434 -
Microorganisms Mar 2021A cold-adapted siphovirus, vB_PagS_AAS23 (AAS23) was isolated in Lithuania using the strain AUR for the phage propagation. The double-stranded DNA genome of AAS23...
A cold-adapted siphovirus, vB_PagS_AAS23 (AAS23) was isolated in Lithuania using the strain AUR for the phage propagation. The double-stranded DNA genome of AAS23 (51,170 bp) contains 92 probable protein encoding genes, and no genes for tRNA. A comparative sequence analysis revealed that 25 of all AAS23 open reading frames (ORFs) code for unique proteins that have no reliable identity to database entries. Based on the phylogenetic analysis, AAS23 has no close relationship to other viruses publicly available to date and represents a single species of the genus within the family . The phage is able to form plaques in bacterial lawns even at 4 °C and demonstrates a depolymerase activity. Thus, the data presented in this study not only provides the information on -infecting bacteriophages, but also offers novel insights into the diversity of cold-adapted viruses and their potential to be used as biocontrol agents.
PubMed: 33807116
DOI: 10.3390/microorganisms9030668 -
Journal of Bacteriology May 2016Cysteine donates sulfur to macromolecules and occurs naturally in many proteins. Because low concentrations of cysteine are cytotoxic, its intracellular concentration is...
UNLABELLED
Cysteine donates sulfur to macromolecules and occurs naturally in many proteins. Because low concentrations of cysteine are cytotoxic, its intracellular concentration is stringently controlled. In bacteria, cysteine biosynthesis is regulated by feedback inhibition of the activities of serine acetyltransferase (SAT) and 3-phosphoglycerate dehydrogenase (3-PGDH) and is also regulated at the transcriptional level by inducing the cysteine regulon using the master regulator CysB. Here, we describe two novel cysteine-inducible systems that regulate the cysteine resistance of Pantoea ananatis, a member of the family Enterobacteriaceae that shows great potential for producing substances useful for biotechnological, medical, and industrial purposes. One locus, designated ccdA(formerly PAJ_0331), encodes a novel cysteine-inducible cysteine desulfhydrase (CD) that degrades cysteine, and its expression is controlled by the transcriptional regulator encoded byccdR(formerly PAJ_0332 orybaO), located just upstream of ccdA The other locus, designated cefA (formerly PAJ_3026), encodes a novel cysteine-inducible cysteine efflux pump that is controlled by the transcriptional regulator cefR(formerly PAJ_3027), located just upstream of cefA To our knowledge, this is the first example where the expression of CD and an efflux pump is regulated in response to cysteine and is directly involved in imparting resistance to excess levels of cysteine. We propose that ccdA and cefA function as safety valves that maintain homeostasis when the intra- or extracellular cysteine concentration fluctuates. Our findings contribute important insights into optimizing the production of cysteine and related biomaterials by P. ananatis
IMPORTANCE
Because of its toxicity, the bacterial intracellular cysteine level is stringently regulated at biosynthesis. This work describes the identification and characterization of two novel cysteine-inducible systems that regulate, through degradation and efflux, the cysteine resistance of Pantoea ananatis, a member of the family Enterobacteriaceae that shows great potential for producing substances useful for industrial purposes. We propose that this novel mechanism for sensing and regulating cysteine levels is a safety valve enabling adaptation to sudden changes in intra- or extracellular cysteine levels in bacteria. Our findings provide important insights into optimizing the production of cysteine and related biomaterials by P. ananatis and also a deep understanding of sulfur/cysteine metabolism and regulation in this plant pathogen and related bacteria.
Topics: Bacterial Proteins; Cystathionine gamma-Lyase; Cysteine; Drug Resistance, Bacterial; Fermentation; Gene Expression Regulation, Bacterial; Homeostasis; Pantoea; Regulatory Elements, Transcriptional; Sulfur; Transcription, Genetic
PubMed: 26883827
DOI: 10.1128/JB.01039-15