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Biomacromolecules , 2 1 , Cultivation temperature modulated the monomer composition and polymer properties of polyhydroxyalkanoate synthesized by Cupriavidus sp. L7L from levulinate as sole carbon source. Gurubasappa G.
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International Journal of Biological Macromolecules , 48 1 , Polymer Degradation and Stability , 95 8 , The deletion was verified using PCR and sequencing.
Pyoverdine was estimated spectrophotometrically as the absorbance at nm. Biofilms were grown statically for 24 h at room temperature. Crystal violet was decanted, and cells were washed five times with water to remove unbound stain. Quantification of three glass tubes, in triplicate experiments, was performed for each strain tested. Three technical replicates were performed for each experiment and each experiment was performed three times.
A markerless in-frame deletion mutant was constructed in the P. The effect of the mutation on bacterial growth was tested by monitoring growth in rich and minimal media. CarA does not influence growth in rich or minimal medium. Growth is represented as OD over time. For each medium, a representative graph of two independent biological experiments is shown. To quantitate this, we measured the pyoverdine production throughout growth in MG medium.
Pyoverdine production was normalized to growth. Pyoverdine production of the wild-type P. A representative graph of two independent biological experiments is shown. As a control, we tested the ability of the wild-type and mutant strain to grow on ammonium as a sole nitrogen source. S1, Supporting Information. Growth of the wild-type P. Mutations in molybdopterin cofactor MoCo synthesis result in the inability to utilize nitrate by either pathway.
Molydenum is also required for xanthine dehydrogenase activity Johnson, Chaudhury and Rajagopalan For most bacteria, MoCo mutants will not grow with hypoxanthine as a sole nitrogen source because the metabolism of the hypoxanthine requires the MoCo-containing enzyme xanthine dehydrogenase. To test if the P. In addition, our data also demonstrate that P. A Biofilm formation was measured by the crystal violet-staining method and expressed as OD The data represent the averages from three independent experiments.
B Diameter of swarming colonies of P. The experiment was repeated two times with similar results. C Diameter of swimming colonies of P. The vertical bars represent the standard deviation of the mean. Although no significant differences in the ability to attach to a glass surface were detected between the strains, we evaluated the strains for their ability to swim and swarm since deficiencies in nitrate assimilation sometimes affect motility, and biofilm formation is often dependent upon motility. It is unknown whether genes that influence utilization of xanthine dehydrogenase or molybdopterin biosynthesis impact growth and virulence in P.
B Representative photographs taken 6 days post inoculation are shown.
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The experiments were repeated three times with similar results. Many members of this family of proteins remain uncharacterized. One example of a protein in this family that has been characterized is PA in P. More recently, Kozmin et al. Additionally, Dahl et al. There is diversity in nitrogen metabolism across Pseudomonas spp.
Pseudomonas syringae is not known to grow anaerobically and lacks the pathways involved in denitrification. Interestingly, a number of the pathways influenced by PA are involved in anaerobic growth; however, many other genes are affected under aerobic conditions.
Ammonia is for many bacteria the preferred source of nitrogen. Ammonium is assimilated into amino acids through use of the glutamate synthase enzymes. Pseudomonas syringae differs from other Pseudomonas because it lacks homologs of gdhA , which encodes glutamate dehydrogenase, amt-3 , an ammonium transporter and a gene for the glutamine synthase type III.
Dahl et al. We hypothesize that the increase in iron—sulfur pools could result in excess iron—sulfur clusters in the P. The importance of nitrate assimilation has been investigated in several plant pathogenic bacteria. Pseudomonas syringae pv syringae Ba uses nitrate as a nitrogen source when cultured on bean leaves Parangan-Smith and Lindow Although a mutant strain lacking NasB did not display reduced growth on bean leaves, a deficiency of growth of the nasB mutant was observed in plant exudates. For Ralstonia solanacearum , a nasA mutant exhibited reduced virulence and delayed stem colonization after soil soak inoculation of tomato plants Dalsing and Allen It is possible that nitrate assimilation is important for the growth and virulence of P.
The xanthine dehydrogenase is a molybdenum-containing flavoprotein that has described roles in a variety of biological processes Wang, Zhang and Xing The importance of xanthine dehydrogenase or MoCo in bacterial plant pathogenesis has not been investigated. MoCo is a tricyclic pterin molecule that is covalently attached to molybdate Williams, Mizrahi and Kana Capturing the redox properties of molybdenum allows enzymes to catalyze many diverse redox reactions important in carbon, nitrogen and sulfur metabolism. Therefore, targeting MoCo biosynthesis may provide an alternative approach to disease management of bacterial plant pathogens.
There is limited information regarding the function of MoCo-dependent enzymes in P. We would like to thank Zhongmeng Bao for construction of the P. Mention of trade names or commercial products in this publication is solely for the purposes of providing specific information and does not imply recommendation or endorsement by the USDA. Oxford University Press is a department of the University of Oxford. It furthers the University's objective of excellence in research, scholarship, and education by publishing worldwide. Sign In or Create an Account.
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Melanie J. Cite Citation. Permissions Icon Permissions. Abstract The plant pathogen Pseudomonas syringae accounts for substantial crop losses and is considered an important agricultural issue. Pseudomonas syringae , PA , nitrate assimilation , molybdopterin , virulence , tomato. Open in new tab Download slide. Influence of the molybdenum cofactor biosynthesis on anaerobic respiration, biofilm formation and motility in Burkholderia thailandensis. Search ADS.