Anastasia Tampakaki

In the present study, in addition to farmyard manure (FYM), cowpea was applied as green manure and faba bean as an intercrop in an organic greenhouse tomato crop, aiming to increase the levels of soil N. Three experiments (E1, E2, E3) were carried out, in which legumes were either noninoculated or inoculated with rhizobia alone or together with plant growth, promoting rhizobacteria. Inoculation of legumes with rhizobia considerably increased N2 fixation in E1 but had no impact on N2 fixation in E2 and E3. In E1, the application of cowpea decreased yield because it imposed a stronger nematode infection as the cowpea plants acted as a good host for Meloidogyne. However, in E2 and E3 the nematode infection was successfully controlled and the legumes significantly increased the tomato yield when inoculated in E2, irrespective of legume inoculation in E3. The total N concentration in the tomato plant tissues was significantly increased by legume application in E2 and E3, but not in E1. T...

Phytopathogenic bacteria affect all kinds of plants and cause several kinds of diseases. However, most plants, in nature, are generally resistant to most pathogens. The range of plant species susceptible to bacterial infection includes the majority of agriculturally important crops. This review highlights current status, insights, and emerging concepts in the taxonomy, pathogenicity mechanisms, lifestyles, pathogen detection, disease diagnosis, and management and progress in the genomics of plant pathogenic bacteria. The study of bacterial pathogen biology has been periodically punctuated by key scientific discoveries, such as the direct transfer of macromolecules (DNA and proteins) across the prokaryote–eukaryote phylogenetic divide. Some of these discoveries served as springboards for breakthrough technologies, such as the genetic engineering of plants, enabled the development of commercial products, and contributed significantly to our understanding the molecular genetic basis of plant–pathogen specificity. The genomic revolution also provided new insights into the evolution of phytobacterial pathogens, the emergence of new pathogen strains/races, host shifts in pathogen populations, and the biology of minimal-size genomes. Future advances in phytopathogenic bacterial genomics and metagenomics will enable the development of more powerful tools for disease epidemiology, diagnosis and management, pathogen quarantine enforcement, and food crop biosecurity.

Type III secretion systems enable plant and animal bacterial pathogens to deliver virulence proteins into the cytosol of eukaryotic host cells, causing a broad spectrum of diseases including bacteremia, septicemia, typhoid fever, and bubonic plague in mammals, and localized lesions, systemic wilting, and blights in plants. In addition, type III secretion systems are also required for biogenesis of the bacterial flagellum. The HrcQ B protein, a component of the secretion apparatus of Pseudomonas syringae with homologues in all type III systems, has a variable N-terminal and a conserved C-terminal domain (HrcQ B -C). Here, we report the crystal structure of HrcQ B -C and show that this domain retains the ability of the full-length protein to interact with other type III components. A 3D analysis of sequence conservation patterns reveals two clusters of residues potentially involved in protein–protein interactions. Based on the analogies between HrcQ B and its flagellum homologues, we ...

With the advent of recombinant DNA techniques, the field of molecular plant pathology witnessed dramatic shifts in the 1970s and 1980s. The new and conventional methodologies of bacterial molecular genetics put bacteria center stage. The discovery in the mid-1980s of the hrp/hrc gene cluster and the subsequent demonstration that it encodes a type III secretion system (T3SS) common to Gram negative bacterial phytopathogens, animal pathogens, and plant symbionts was a landmark in molecular plant pathology. Today, T3SS has earned a central role in our understanding of many fundamental aspects of bacterium-plant interactions and has contributed the important concept of interkingdom transfer of effector proteins determining race-cultivar specificity in plant-bacterium pathosystems. Recent developments in genomics, proteomics, and structural biology enable detailed and comprehensive insights into the functional architecture, evolutionary origin, and distribution of T3SS among bacterial pa...

The C-terminal domain of the HrcQ(B) protein from the Hrp secretion system of the plant pathogenic bacterium Pseudomonas syringae pv. phaseolicola has been crystallized from MPD using the hanging-drop vapour-diffusion method. The crystals belong to space group P2(1), with unit-cell parameters a = 51.6, b = 27.3, c = 97.2 A and beta = 99.8 degrees. A complete native data set extending to 3.0 A resolution was collected from a single cryoprotected crystal. The crystal solvent content and calculation of self-rotation functions showing non-crystallographic twofold symmetry axes are consistent with the presence of an oligomeric assembly in the asymmetric unit.

Information about the availability of soil mineral nitrogen (N) in organic greenhouse tomatoes after the application of mobile green manure (MGM), and its impact on plant nutrient status and yield is scarce. Considering this knowledge gap, the effects of legume biomass from faba beans that are cultivated outdoors (FAB), or from feed-grade alfalfa pellets at two different doses (AAL = 330 g m−2; AAH = 660 g m−2) that were applied as MGM on the nutrition and yield of an organic greenhouse crop of tomatoes were evaluated. All of the MGM treatments increased the mineral N concentrations in the soil throughout the cropping period, and the total N concentration in tomato leaves when compared to the untreated control. FAB and AAH treatments had a stronger impact than AAL in all of the measured parameters. In addition, AAL, AAH, and FAB treatments increased the yield compared to the control by 19%, 33%, and 36%, respectively. The application of MGM, either as faba bean fresh biomass or as a...

The ability of the Pseudomonas syringae pv. phaseolicola harpin (HrpZPsph) to elicit hypersensitive response was investigated in three Nicotiana genotypes. The hrpZPsph gene was placed under chemical regulation (tetracycline induction) in TetR+ Nicotiana tabacum cv. Wisconsin 38 (W38) or was transiently expressed in N. benthamiana following infection with a PVX-derived vector and in three Nicotiana genotypes by agroinfiltration. The constructs were designed to express either the canonical form of harpin (HrpZPsph) or an N-terminally extended version of the protein carrying the signal peptide portion of the tobacco pathogenesis-related protein PR1a (SP-HrpZPsph). Stable transformants of N. tabacum cv. W38 did not develop necrosis upon induction with tetracycline, probably as a result of insufficient harpin accumulation. In contrast, N. benthamiana plants infected with the PVX constructs produced high concentrations of harpin in biologically active form, but only those expressing the secretable form of harpin developed necrotic symptoms. These symptoms were less severe than those caused by PVX::avrPto; however, they were accompanied by induction of hsr203J, a hypersensitive response-specific gene transcript. These results suggest that the plant cellular receptor(s) for harpin is extracellular.

The soil nitrogen-fixing bacterium Azotobacter vinelandii possesses two cyclophilins, comprising putative cytoplasmic and periplasmic isoforms, designated as AvPPIB and AvPPIA, respectively. Both recombinant cyclophilins have been purified and their peptidyl-prolyl cis/trans isomerase activity against Suc-Ala-Xaa-Pro-Phe-pNA synthetic peptides has been characterized. The substrate specificity of both cyclophilins is typical for bacterial cyclophilins, with Suc-Ala-Ala-Pro-Phe-pNA being the most rapidly catalyzed substrate. The cytoplasmic cyclophilin also displays a chaperone function in the citrate synthase thermal aggregation assay. Using real-time quantitative RT-PCR, we demonstrate that AvppiB is expressed under various physiological and growth conditions, mainly upregulated by acetate and downregulated by the stationary growth state, while AvppiA shows a tendency for downregulation under the tested conditions. Further, we identified chaperone protein dnaK and UDP-2, 3-diacylglucosamine hydrolase lpxH as probable interacting partners of AvPPIB and we demonstrate their physical interaction by coexpression studies. An increase in AvPPIB PPIase activity in the presence of AvdnaK and a decrease in the presence of AvlpxH further confirms each interaction. However, the PPIase activity does not seem to be essential for those interactions since AvPPIB active site mutants still interact with dnaK and lpxH, while their minor PPIase activity cannot be modulated by the interaction.