doi: 10.1094/MPMI-09-11-0253, Withnall, R., Chowdhry, B. Biol. nov. Int. 18, 385–396. 134, 2449–2455. Finally we demonstrate that the ΔcrtB mutant shows reduced colonization of plant roots. The production of indoles was measured using a colorimetric assay. As predicted, the ΔcrtB mutant no longer produced a yellow pigment and the colonies appeared white (Figure 4B). Indeed, the mutant was defective in both surface attachment and pellicle formation. doi: 10.1111/j.1751-1097.1991.tb01989.x, Kamilova, F., Kravchenko, L. V., Shaposhnikov, A. I., Azarova, T., Makarova, N., and Lugtenberg, B. 31, 425–448. 31, 447–460. To compare growth rates of Pantoea sp. 78, 6859–6865. The United States Government retains and the publisher, by accepting the article for publication, acknowledges that the United States Government retains a non-exclusive, paid-up, irrevocable, world-wide license to publish or reproduce the published form of this manuscript, or allow others to do so, for United States Government purposes. 60, 2430–2440. Raman spectroscopic study of the heterogeneity of microcolonies of a pigmented bacterium. Indole-3-acetic acid production was measured by a colorimetric assay as previously described (Tang and Bonner, 1947). JM-F, AB, and MD developed the experimental plan. Raman spectra of carotenoids in natural products. MBDS Solvent: an improved method for assessment of biofilms. Cell viability was measured using the Bac-Titer Glo assay and plotted as a percentage relative to the untreated control, measured as 100%. 1998 to the genus as Pantoea cypripedii comb. A., Penna, C., Ruiz, O. Appl. YR343 and P. ananatis LMG 20103 (De Maayer et al., 2010) indicated that the amino acid sequences of each gene from Pantoea sp. YR343 grown in liquid LB, R2A, and M9 media. Boca Raton: CRC Press. Due to the observation that the ΔcrtB mutant is impaired in biofilm formation and IAA production, we speculated that this mutant may also be affected in root colonization. FIGURE 5. Environ. PCR products were ligated into pK18mobsacB (Schafer et al., 1994) and the resulting plasmid was verified by restriction digests and transformed into Pantoea sp. YR343, a non-pathogenic bacterial strain isolated from the rhizosphere of poplar, which was found to be a robust colonizer of plant roots. We next analyzed the ability of Pantoea sp. In P. ananatis, it was found that deletion of the phytoene synthase gene, crtB, resulted in the loss of yellow pigment and increased sensitivity to environmental stress factors (Mohammadi et al., 2012). Genetic organization of the cellulose synthase operon in Acetobacter xylinum. In this case, Pantoea sp. (C) Colony morphology of Pantoea sp. nov., Pantoea rwandensis sp. (1994). YR343 produced a yellow pigment under all growth conditions tested; however, it was the most apparent when cells were grown to stationary phase in LB medium. Molecular biology of cellulose production in bacteria. Int. YR343 Raman spectrum. La formation doit permettre la réalisation d’activités centrées sur l’analyse des solutions existantes, ainsi que sur la conception et la définition de produits industriels. YR343 to establish a strong interaction with the plant root, perhaps due to defects in IAA production or in biofilm production. Mol. 70, 249–256. MBio 6:e01251. (1987). doi: 10.1351/pac198557050785, Mohammadi, M., Burbank, L., and Roper, M. C. (2012). doi: 10.1590/S1415-47572012000600020, Brady, C. L., Cleenwerck, I., van der Westhuizen, L., Venter, S. N., Coutinho, T. A., and De Vos, P. (2012). Our genome comparisons, however, suggest that Pantoea sp. doi: 10.1016/S0378-1119(96)00423-4, Dussault, D., Caillet, S., Le Tien, C., and Lacroix, M. (2008). Mol. (A) Comparison of biofilm formation between wild type Pantoea sp. Many soil-dwelling and plant-associated bacteria, including Pantoea spp., produce carotenoids, which are pigment molecules found in the cellular membrane (Dussault et al., 2008; Mohammadi et al., 2012). YR343 (data not shown). Imaging of root colonization was performed by staining root tissue with 5 μM Syto61 (Life Technologies) to stain all bacterial cells that were attached to the root. 7:491. doi: 10.3389/fmicb.2016.00491. As predicted from these analyses, loss of phytoene synthase activity by deletion of the crtB gene, resulted in a strain that was unable to produce any detectable carotenoids and showed increased sensitivity to reactive oxygen species. YR343 using P. deltoides, T. aestivum (wheat), or A. thaliana as plant hosts. Partial nucleotide sequences from atpD, infB, gyrB, and rpoB were used for multi locus sequence analysis. Identification and characterization of a carotenoid mutant in Pantoea sp. New functional assignment of the carotenogenic genes crtB and crtE with constructs of these genes from Erwinia species. Appl. Biopolymers 77, 212–221. YR343 in this paper. Med. doi: 10.1016/S0923-2508(02)01316-5, Ruppel, S., Hecht-Buchholz, C., Remus, R., Ortmann, U., and Schmelzer, R. (1992). Pantoea sp. YR343 was grown on LB supplemented with 0.4% glycerol (Figure 2E). YR343 and measurement of colonization. YR343 readily colonizes plant roots and does not appear to be pathogenic when applied to the leaves or roots of selected plant hosts. 100, 938–945. Received: 19 January 2016; Accepted: 24 March 2016;Published: 18 April 2016. Landrum, J. T. (2009). VII. Here we describe Pantoea sp. Syst. Genomic comparisons indicate the presence of two cellulose synthase operons in other Pantoea spp. Cells that only fluoresced red were assumed to be ΔcrtB mutants, and cells that fluoresced red and green were assumed to be wild type cells expressing GFP. Lett. Biological role of pigment production for the bacterial phytopathogen Pantoea stewartii subsp. Plant Dis. Oxidative burst: an early plant response to pathogen infection. (2002). By this method, we observed a steady decrease in the spectral peak intensity corresponding to zeaxanthin (Figure 4F) which correlates to the loss of cell viability. Phylogenetic tree of Pantoea sp. Plants treated with sterile R2A medium were used as controls for background contamination. Mol. 132, 3407–3414. As predicted, the ΔcrtB mutant is defective in carotenoid production, and shows increased sensitivity to oxidative stress. doi: 10.1007/s13213-010-0117–111, Herrera, C. M., Koutsoudis, M. D., Wang, X., and von Bodman, S. B. Appl. (B) Corresponding to the pictures in (A) are images of roots taken with confocal microscopy. Plant Microbe Interact. Loss of carotenoids affects growth and phytohormone production. Other potential pathways to IAA production include the indole-acetamide pathway, although it is difficult to determine by genome comparisons whether this pathway is complete in Pantoea sp. Biotechnol. 77, 5934–5944. J. The strong Raman signal in wild type Pantoea sp. GM01; Brown et al., 2012) within a monophyletic Pantoea group (90%) and forms a unique, well supported (100%) group, basal to the recently described P. rwandensis and P. rodasii, that are known to form lesions on leaves of plantation-grown Eucalyptus trees (Brady et al., 2012) (Figure 1). Genomic analyses of Pantoea sp. (2008). doi: 10.1094/MPMI-18–0385, Wasim, M., Bible, A. N., Xie, Z., and Alexandre, G. (2009). 69, 253–257. YR343 also appeared to be highly mucoid, particularly when grown on R2A medium. With the advent of fingerprinting techniques, many traditional concepts regarding transmission, infectivity, or pathogenicity of mycobacterial bacilli have been revisited, and their conventional interpretations have been challenged. Pseudomonas fluorescens induces strain-dependent and strain-independent host plant responses in defense networks, primary metabolism, photosynthesis, and fitness. (2001). gypsophilae. Indeed, we observed a nearly threefold decrease in indole production from 4.35 ± 0.20 μg/ml indoles in wild type cells compared to 1.52 ± 0.02 μg/ml indoles in ΔcrtB. Pantoea sp. Tarifs Rentrée 2012. Biochim. 1973 emend. Philos. doi: 10.1111/j.1365-2672.2006.02843.x, Foote, C. S., and Denny, R. W. (1968). (2013). Fluorescent strains of Pantoea sp. (2015). Krinsky, N. I. Bioessays 28, 1091–1101. Flagella-driven chemotaxis towards exudate components is an important trait for tomato root colonization by Pseudomonas fluorescens. Mol. The decrease in Raman signal intensity was apparent even at H2O2 concentrations that did not result in decreased cell viability, consistent with the protective role of carotenoids in the presence of free oxygen radicals (Krinsky, 1989, 1993; Hirayama et al., 1994; Edge et al., 1997). Genet. Swimming and swarming motility was examined on LB containing 0.3% w/v agar or 0.6% w/v agar supplemented with 0.4% w/v glucose or 0.4% v/v glycerol, based on previous studies (Herrera et al., 2008). Plant Microbe Interact. It is thought that P. agglomerans promotes plant growth by enhancing root architecture, which increases the amount of minerals and water that can be taken up by the plant (Sergeeva et al., 2007). YR343 was isolated from a healthy poplar tree, its close phylogenetic relationship to Pantoea strains found to be the causal agents of leaf blight in Eucalyptus prompted us to test the effect of Pantoea sp. Pantoea sp. doi: 10.1016/j.syapm.2008.09.004, Brady, C. L., Cleenwerck, I., Venter, S. N., Engelbeen, K., De Vos, P., and Coutinho, T. A. Many beneficial microbes have developed a number of strategies for overcoming such plant defense mechanisms (Zamioudis and Pieterse, 2012). Chamberlain, N. R., Mehrtens, B. G., Xiong, Z., Kapral, F. A., Boardman, J. L., and Rearick, J. I. l standing pro coach who will re ceive a trophy at the Touchdown j Club’s annual banquet January 8 J at the Hotel Statler. Carotenoids and bacterial photosynthesis: the story so far. Métier. Appl. Microbiol. The carotenoid biosynthesis pathway has been well characterized in Pantoea and consists of six enzymes: geranylgeranyl diphosphate (GGPP) synthase CrtE, phytoene synthase CrtB, phytoene desaturase CrtI, lycopene cyclase CrtY, β-carotene hydroxylase CrtZ, and the zeaxanthin glucosyltransferase CrtX (To et al., 1994; Sedkova et al., 2005). Phytopathology 100, 1330–1339. (1947). An overnight culture was diluted 1:100 into either LB, R2A, or M9 medium supplemented with 0.4% w/v glucose and grown statically in a 96-well plate covered with breatheable tape in place of the lid (Breathe-EASIER, Diversified Biotech) at 28°C for 72 h. After 72 h, adherent cells were stained with 0.1% w/v crystal violet stain, then the crystal violet associated with biofilms was dissolved using a modified solution which contained 10% w/v SDS dissolved in 80% v/v ethanol (Tram et al., 2013). stewartii exhibits surface motility, which is a critical aspect of Stewart’s wilt disease development on maize. Although Pantoea sp. YR343 in the soil, which would therefore influence its ability to associate with a plant root. Young, G. Britton and R. J. Cogdell (Berlin: Springer). 15, 473–497. From this perspective, the decrease in IAA production by the ΔcrtB mutant may be a consequence of changes in membrane fluidity or organization. Wild type and mutant cells were grown to the same optical density and the supernatants were measured for the presence of IAA. Conversely, little to no pigment was extracted from the ΔcrtB mutant (Figure 4C). Motility, directed by chemotaxis, is an important means by which bacteria may avoid adverse conditions, while detecting and colonizing roots within the soil environment (Bashan, 1986; de Weert et al., 2002; Somers et al., 2004). Syringae (Braun-Kiewnick et al., 2000). (1998). Norm Van Brocklin, Los An- j geles Rams’ back, will be honored ; as the outstanding pro player. Among the organisms isolated from the Populus rhizosphere was the γ-proteobacterium Pantoea sp. Evaluation of plant growth promoting and colonization ability of endophytic diazotrophs from deep water rice. Chem. Graph represents the range of absorbances between 400 and 500 nm measured from one of two replicates. Gene 145, 69–73. YR343 and ΔcrtB. (Right) Cellulose production in Pantoea sp. Cultures were imaged using a Zeiss LSM 710 laser scanning confocal microscope and images were processed using Zen software (Zeiss). This process results in the selective enhancement, by factors of up to 106, of the Raman intensities of bands coupled to the electronic transition of the chromophore, which can be advantageous in the analysis of complex systems (Robert, 1990). 35, 1044–1051. YR343 displays a rod-shaped morphology with cells averaging approximately 2 μm in length (Figure 2D). For soil samples, 1 g of soil was mixed with 4 ml of PBS, then vortexed and plated similarly. After 18 h, the cells had moved from the center to the edges of the plate, consistent with swimming motility behavior. (2009). Functional microdomains in bacterial membranes. YR343 in the presence of 0.4% glycerol, as compared to medium containing 0.4% glucose. This research was sponsored by the Genomic Science Program, U.S. Department of Energy, Office of Science, Biological and Environmental Research, as part of the Plant Microbe Interfaces Scientific Focus Area (http://pmi.ornl.gov). doi: 10.1146/annurev.arplant.48.1.251. Since only two isolates are available from Populus in the Eastern USA at present, it is unclear at this point whether these isolates represent a single or multiple new species. The tryptophan-dependent production of IAA was confirmed by GC-MS analyses and measured at approximately 0.5 μg/ml. doi: 10.1128/AEM.05255-11. Production of indole-3-acetic acid, aromatic amino acid aminotransferase activities and plant growth promotion by Pantoea agglomerans rhizosphere isolates. YR343 was isolated from the rhizosphere of a healthy Populus deltoides tree in North Carolina and is a robust colonizer of plant roots. “Influence of growth promoting bacteria from Uzbekistan and Germany on the growth and nutrient uptake of cotton and wheat on different soils,” in Plant Nutrition: Food Security and Sustainability of Agro-Ecosystems Through Basic and Applied Research Developments in plant and soil sciences, eds W. Horst, M. K. Schenk, A. Bürkert, N. Claassen, H. Flessa, W. B. Frommer, et al. syringae, the causal agent of basal kernel blight of barley, by antagonistic Pantoea agglomerans. 71, 8141–8146. Analysis of the gene cluster encoding carotenoid biosynthesis in Erwinia herbicola Eho13. doi: 10.1094/MPMI-21-10-1359, Hirayama, O., Nakamura, K., Hamada, S., and Kobayasi, Y. Microbiol., 18 April 2016 As determined by phylogenetic analysis, Pantoea sp. Bliv medlem af Facebook, og få kontakt med Anaïs Rouvière og andre, du måske kender. YR343 could produce IAA, which is synthesized from the amino acid tryptophan (Patten and Glick, 1996). Plant Biol. YR343 was isolated from the rhizosphere of a native Populus deltoides tree growing in the Yadkin River region of North Carolina (Shakya et al., 2013) and a draft genome sequence was obtained (Brown et al., 2012). YR343 growing on LB agar plates produced colonies that were round, smooth, and produced a yellow pigment (Figure 2C). Ultimately, the phenotypes associated with the loss of carotenoids in Pantoea sp. doi: 10.1111/j.1472-765X.2008.02410.x, Edge, R., McGarvey, D. J., and Truscott, T. G. (1997). doi: 10.1094/Phyto.03.10.0097. 192, 2936–2937. Aucun avis n'a été posté sur Lycée Rouvière. Each Raman spectrum recorded was an accumulation of 100 spectra acquired with integration time of 0.5 s each at 5 mW incident laser power. Roots were harvested as described above for Populus and analyzed for CFUs per gram of root material. Biological Control of Pseudomonas syringae pv. Jil Benedetto. YR343 exopolysaccharides was detected by inoculating 200 μl of an overnight culture into a glass-bottom dish containing 5 ml of R2A and grown statically at 25°C for 72 h. After incubation, the culture was stained with a solution of Calcofluor White (5 μg ml-1) (Sigma–Aldrich), and 5 μM SYTO61 (Life Technologies). In this experiment, the mutant cells tended to settle at the bottom of the tube rather than form a biofilm at the air-liquid interface like wild type cells. Plant. 28, 991–998. doi: 10.1094/Pdis.2000.84.9.973, Barret, M., Morrissey, J. P., and O’Gara, F. (2011). Rev. YR343 in LB, R2A, or M9 media, overnight cultures were diluted 1:100 into fresh medium and 200 μl was loaded into 12 wells on a honeycomb plate and placed into a Bioscreen C Reader System at room temperature with shaking overnight. 13, 561–587. Rooted cuttings were planted in sterile clay soil inoculated with Pantoea sp. Soc. Modulation of host immunity by beneficial microbes. Swarming motililty has been associated with virulence in the plant pathogen, P. stewartii, which colonizes the plant xylem, blocking flow and causing wilting in its plant hosts, including corn (Herrera et al., 2008). Regulation of membrane lipid fluidity in Acholeplasma laidlawii: effect of carotenoid pigment content. Hopanoid biosynthesis and function in bacteria. Sci. Shoot tips were then washed three times in sterile water before inoculation into MS medium (per 1 l of medium: 4.43 g MS salts, 0.5 g MES hydrate, 30 g sucrose, 5 g activated charcoal, 1.5 g Gelrite and 1 ml plant preservative mixture (PPM). The bacteria were aggregated into what appeared to be biofilms on the surface of the roots and along the root hairs. doi: 10.1099/Ijs.0.017301–17300, Brady, C. L., Venter, S. N., Cleenwerck, I., Engelbeen, K., Vancanneyt, M., Swings, J., et al. AB, SF, DP, and JM-F performed phenotypic characterization and mutant analyses; CS and TC performed phylogenetic analyses, NE and TT performed GC-MS analyses, RM, SP, and PB performed Raman spectroscopy analyses, and AB, SJ, and DW conducted pathogenicity analyses. Sequences from Populus rhizosphere isolates GM01, YR343 (Brown et al., 2012) and three other unidentified isolates for which data was available from genome sequencing efforts on IMG were aligned using the Translation alignment tool within GeneiousTM (v6.0.4-www.geneious.com) with 94 total representatives of Pantoea and related taxa of Erwinia, Tatumella and other Enterobacteriaceae for further phylogenetic analysis with Chronobacter sakazakii as an outgroup. Plant Soil 145, 261–273. Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. (2001). Pantoea: insights into a highly versatile and diverse genus within the Enterobacteriaceae. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). To enable detection of Panteoa sp. Approximately 500 base pairs from the 5′ end of the 16S rRNA gene were amplified using universal primers (Weston et al., 2012). Natural infestation of onion seed by Pantoea ananatis, causal agent of center rot. Consistent with this data, we observed YR343-pGFP cells attached to P. deltoides roots using confocal microscopy, whereas no bacteria were detected in the un-inoculated control plants (Figure 3C). (1991). Microbiol. Pantoea sp. *Correspondence: Jennifer L. Morrell-Falvey, firstname.lastname@example.org, Front. (B) Comparison of indole-3-acetic acid production in the wild type Pantoea sp. Afterward, seeds were washed briefly in 70% ethanol and rinsed several times in sterile water. Based on phylogeny, Pantoea sp. Environ. Pantoea sp. doi: 10.1016/S0734-9750(99)00014-2, Romling, U. Microbiol. Genome sequence of Pantoea ananatis LMG20103, the causative agent of Eucalyptus blight and dieback. Moreover, we find that the ΔcrtB mutant is impaired in biofilm formation and production of IAA. Briefly, roots were weighed, washed by vortexing at least 30 s with 3 ml of PBS and a few small glass beads and dilutions of the wash solution was plated on R2A to measure colony forming units (CFU). doi: 10.1007/s11104-008-9568-6, Robert, B. Interestingly, IAA is produced in some pathogenic species of Pantoea that induce tumor or gall formation via the tryptamine pathway (Manulis et al., 1998). Microbiol. Resonance Raman spectroscopy of carotenoids and carotenoid-containing systems. Co-cultures of Arabidopsis seedlings and Pantoea sp. YR343, a Bacteria Isolated from the Rhizosphere of Populus deltoides, Is Defective in Root Colonization. The enzymatic inactivation of indoleacetic acid; some characteristics of the enzyme contained in pea seedlings. 17, 319–339. doi: 10.1080/10408410490468786, Spaepen, S., Vanderleyden, J., and Remans, R. (2007). 7, 3319–3328. 28, 449–461. (Berlin: Springer), 674–675. (2010). Plant Dis. Phylogeny and identification of Pantoea species associated with plants, humans and the natural environment based on multilocus sequence analysis (MLSA). YR343. Plant Physiol. FIGURE 2. It will be interesting to perform comparative genomic analyses on new and related Pantoea strains as they become available to resolve these systematics issues and to determine whether any host-specificity or pathogenicity-related factors can be identified. Métier. YR343 grown in R2A medium prior to planting the newly rooted Poplar plant. nov., and transfer of Pectobacterium cypripedii (Hori 1911) Brenner et al. Moreover, the carotenoid biosynthesis genes in Pantoea sp. Plant Microbe Interact. YR343 and ΔcrtB in M9 minimal medium. 24, 1893–1902. YR343 is shown by a zone of clearing surrounding the colony. Evol. For each sample, individual Raman spectra were collected at 15 randomly selected spatial locations and averaged to give the representative spectrum for that sample. Nutr. Quenching by beta-carotene. Mol. 47, 208–213. Another possibility is that the loss of carotenoids affects membrane organization and impacts the signaling pathways involved in plant recognition. Potential of NIR-FT-Raman spectroscopy in natural carotenoid analysis. Initial BLAST (16S and gyrB) and Ribosomal Database Project 16S sequence comparisons suggested a strong affiliation of strainYR343 with Pantoea spp. Reactive oxygen species signaling in response to pathogens. Arch. Acceptez-vous de recevoir des notifications d'orientation ? Photothèque. YR343 also allowed us to follow the effect of H2O2 on carotenoids. Bacterial biosynthesis of indole-3-acetic acid. FIGURE 6. Whether these cellulose gene clusters are differentially regulated and/or produce distinct cellulose synthase complexes has yet to be determined. doi: 10.1094/mpmi-06-11-0179. doi: 10.1002/bies.20493, Goodwin, J. R., Hafner, L. M., and Fredericks, P. M. (2006). Likewise, we also observed changes in biofilm formation in the ΔcrtB mutant. Maximum likelihood analysis was conducted using PhyML v3.0, (Guindon et al., 2010) as implemented within GeneiousTM. Fine roots and associated rhizophere samples were harvested from native Populus deltoides trees at the Yadkin River in North Carolina in May 2011 and bacteria were isolated from both the endosphere and rhizosphere. doi: 10.1099/00221287-134-9-2449, Hayat, R., Ali, S., Amara, U., Khalid, R., and Ahmed, I. Loss of carotenoids affects biofilm formation and root colonization. doi: 10.1016/S0168-1656(01)00333-9, Walcott, R. R., Gitaitis, R. D., Castro, A. C., Sanders, F. H., and Diaz-Perez, J. C. (2002). Strains of P. agglomerans have also been shown to promote plant growth in wheat, rice, and cotton (Ruppel et al., 1992; Amellal et al., 1998; Egamberdiyeva and Höflich, 2001; Verma et al., 2001; Feng et al., 2006). Biophys. The bacterial superoxide dismutase and glutathione reductase are crucial for endophytic colonization of rice roots by Gluconacetobacter diazotrophicus PAL5. J. Biotechnol. While the catalytic proteins, encoded by bscA and bscB, are conserved in both types of cellulose synthesis gene clusters, there are genes unique to each operon for which the functions are not well known (Romling, 2002). The results of multi locus sequence analysis (MLSA) using partial nucleotide sequences from atpD, infB, gyrB, and rpoB (Brady et al., 2012) revealed that Pantoea sp. All authors contributed intellectual input and data analyses and assisted in manuscript preparation. Adv. Free Radic. Due to the selective enhancement afforded by resonance Raman scattering, molecular vibrations arising from other cellular components, such as DNA and proteins, are represented by relatively weak bands compared to the carotenoid bands in the Pantoea sp. YR343 on cultured poplar cuttings. Copyright © 2016 Bible, Fletcher, Pelletier, Schadt, Jawdy, Weston, Engle, Tschaplinski, Masyuko, Polisetti, Bohn, Coutinho, Doktycz and Morrell-Falvey. Carotenoids as modulators of lipid membrane physical properties. A representative Raman spectrum of wild type Pantoea sp. YR343 readily forms biofilms on abiotic and biotic surfaces. We then tested the protective role of this carotenoid under oxidative stress created by exposure to hydrogen peroxide. doi: 10.1007/s00253-007-0909–909, Raaijmakers, J. M., Paulitz, T. C., Steinberg, C., Alabouvette, C., and Moenne-Loccoz, Y. Lett. Co-cultured seedlings were harvested and mounted on slides prior to imaging with a Zeiss LSM710 confocal laser scanning microscope. 38, 750–759. Top left image shows a group of motile cells outside of the plant. Microbiol. YR343 and ΔcrtB mutant as described (Mohammadi et al., 2012). 10 μl of the cell suspension was pipetted onto a clean gold-coated silicon wafer and dried for Raman analysis. YR343. (C) Wheat root colonization assay by wild type Pantoea sp. This was further confirmed by analyzing the spectroscopic profiles of pigments extracted from wild type and ΔcrtB mutant cells. Twenty-one genome sequences from Pseudomonas species and 19 genome sequences from diverse bacteria isolated from the rhizosphere and endosphere of Populus deltoides. After 7 days, 4 – 6 A. thaliana seedlings of equivalent root lengths were transferred to new MS agar plates containing 0.25% w/v sucrose and grown for an additional 7 days. doi: 10.1002/jrs.2493, de Weert, S., Vermeiren, H., Mulders, I. H., Kuiper, I., Hendrickx, N., Bloemberg, G. V., et al. Biochim. doi: 10.1111/j.1744-7909.2010.00933.x, O’Toole, G. A., and Kolter, R. (1998). YR343 is detected by GFP fluorescence (green) and plant roots are detected using autofluorescence (red). Cultures were also grown in standard LB, M9, TY (per 1 l, 10 g tryptone, 5 g yeast extract), or SOBG medium (per 1 l, 20 g tryptone, 5 g yeast extract, 0.5 g NaCl, 2.4 g MgSO4, 0.186 g KCl, 50 ml of 40% v/v glycerol). Acta 1017, 99–111. Acta 895, 63–79. (2010). B 41, 189–200. Liaaen-Jensen, S., and Andrewes, A. G. (1972). YR343 for 10 days prior to imaging. Plant Soil 321, 341–361. (2006). La poursuite d’études des bacheliers professionnels (animateur : Emmanuel Quenson, Université d’Évry, Centre Pierre Naville)- Claire Lemêtre (Université Paris 8, CIRCEFT-ESCOL), Juliette Mengneau (Université de Nantes, CENS) \u0026 Sophie Orange (Université de Nantes, CENS), « La fac, on me dit que c'est possible mais que c'est pas faisable ». 48, 251–275. The pigments extracted from wild type cells showed a spectra consistent with zeaxanthin as has been described for other Pantoea strains (Hundle et al., 1991; Mohammadi et al., 2012). (2000). 21, 1359–1370. (1974). YR343 was preferentially associated with the plant as indicated by 3.5 × 106 CFU g-1 of root, compared to 3.0 × 102 CFU g-1 of soil. In this case, we found a mixture of wild type and ΔcrtB mutant cells (Figure 6D), most of which appeared to be motile (data not shown). YR343 was more than 50% identical to those from P. ananatis LMG20103. Colonies lacking yellow pigmentation were screened by PCR to verify that the crtB gene was deleted. Environ. Interestingly, this phenotype was the direct opposite of that shown for P. stewartii (Herrera et al., 2008), suggesting a possible difference in metabolism and preferred growth environment for Pantoea sp. Overnight cultures of wild type Pantoea sp. In other organisms, such as Bacillus subtilis, it was shown that zaragozic acid, an inhibitor of squalene synthesis, could reduce the ability of B. subtilis to form biofilms (Lopez and Kolter, 2010). (2002). J. Appl. The presence of cellulose in Pantoea sp. doi: 10.1046/j.1365-2958.1998.00797.x, Patten, C. L., and Glick, B. R. (1996). Feng, Y., Shen, D., and Song, W. (2006). In all images, Pantoea sp. A multifactor analysis of fungal and bacterial community structure in the root microbiome of mature Populus deltoides trees. A., Cassan, F. D., et al. YR343, indicating that this species is capable of phosphate solubilization (Figure 2B). Microbiol. Bacterial blight and dieback of Eucalyptus species, hybrids, and clones in South Africa. The complex interactions between plants and their microbiome can have a profound effect on the health and productivity of the plant host. Reactive oxygen species during plant-microorganism early interactions. Plant Physiol. 7, 617–635. doi: 10.1111/j.1399-3054.1962.tb08052.x, Nanda, A. K., Andrio, E., Marino, D., Pauly, N., and Dunand, C. (2010). Carotenoids’ influence on radiotolerance of Pantoea agglomerans, a plant pathogen. Pantoea sp. (A) Growth of wild type Pantoea sp. Microbiol. doi: 10.1071/FP11173, Doke, N., Miura, Y., Sanchez, L. M., Park, H. J., Noritake, T., Yoshioka, H., et al. doi: 10.1016/S1011-1344(97)00092-4. In these analyses, we see a steady decrease in the Raman spectra associated with carotenoids as the cells are exposed to increasing concentrations of H2O2. Likewise, ΔcrtB mutant cells were also impaired in pellicle formation (Figure 6B). Similarly, many microbes produce carotenoids, which play a vital role in the survival of cells under harsh conditions, such as oxidative stress, extremes in pH, and resistance to toxins (Liaaen-Jensen and Andrewes, 1972; Krinsky, 1989; Kulkarni et al., 2015). Intégrer l'école' Lycée Rouvière ? Plant Microbe Interact. For the second experiment, we used a culture with 1 × 108 cells per ml of Pantoea sp. (1997). Bottom image shows a three-dimensional view of a colonized wheat root. In addition, carotenoids can modulate membrane fluidity and may play a role in the formation of membrane domains (Huang and Haug, 1974; Chamberlain et al., 1991; Gruszecki, 1999; Gruszecki and Strzalka, 2005; Landrum, 2009; Lopez and Kolter, 2010). GM01 which was isolated from the rhizosphere of a P. deltoides tree in Tennessee. PMI39_03412 encodes a product with 60% amino acid identity and 74% amino acid similarity to P. ananatis CrtE; PMI39_03408 encodes a product with 66% amino acid identity and 76% amino acid similarity to P. ananatis CrtB; PMI39_03409 encodes a product with 81% amino acid identity and 87% amino acid similarity to P. ananatis CrtI; PMI39_03410 encodes a product with 58% amino acid identity and 74% amino acid similarity to P. ananatis CrtY; PMI39_0340 encodes a product with 84% amino acid identity and 92% amino acid similarity to P. ananatis CrtZ, and PMI39_03411 encodes a product with 53% amino acid identity and 61% amino acid similarity to P. ananatis CrtX.
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