Performance of a cryo-cooled crystal monochromator illuminated by hard X-rays with MHz repetition rate at the European X-ray Free-Electron Laser.

Due to the high intensity and MHz repetition rate of photon pulses generated by the European X-ray Free-Electron Laser, the heat load on silicon crystal monochromators can become large and prevent ideal transmission in Bragg diffraction geometry due to crystal deformation. Here, we present experimental data illustrating how heat load affects the performance of a cryogenically cooled monochromator under such conditions. The measurements are in good agreement with a depth-uniform model of X-ray dynamical diffraction taking beam absorption and heat deformation of the crystals into account.

Materials Imaging and Dynamics (MID) instrument at the European X-ray Free-Electron Laser Facility.

The Materials Imaging and Dynamics (MID) instrument at the European X-ray Free-Electron Laser (EuXFEL) facility is described. EuXFEL is the first hard X-ray free-electron laser operating in the MHz repetition range which provides novel science opportunities. The aim of MID is to enable studies of nano-structured materials, liquids, and soft- and hard-condensed matter using the bright X-ray beams generated by EuXFEL. Particular emphasis is on studies of structure and dynamics in materials by coherent scattering and imaging using hard X-rays. Commission of MID started at the end of 2018 and first experiments were performed in 2019.

First Report of the Root-Knot Nematode Meloidogyne hapla Parasitizing Roses in Ethiopia.

Meloidogyne hapla is one of the most damaging plant-parasitic nematodes in temperate regions. This nematode has a wide host range with more than 500 plant taxa including roses. In Ethiopia, rose production has developed over the past 10 years to the second most important export market after coffee. Considering the high damage potential of M. hapla, infestation of roses in Ethiopia with this nematode could result in major economic losses. Therefore, awareness of this nematode species is extremely important. During two surveys conducted in August 2011 and April 2012, M. hapla was detected in soil samples from six out of nine rose producing farms located in the districts of Ziway, Holleta, Sebeta, and Menagesha. At infested farms, rose plants appeared stunted and less productive and often showed symptoms of chlorosis and wilting. Identification was based on morphological and morphometrical characters of females, males, and second-stage juveniles, which were all within the range of variability known for this species (4). Shape of juvenile stylet knobs, shape of male head, and perennial pattern of the females with characteristic punctuations between the anus and tail terminus were also typical for M. hapla. The morphological identification was confirmed by sequence analysis of the D2-D3 expansion segment of the 28S rDNA gene following amplification with the primers D2A (5'-ACAAGTACCGTGAGGGAAAGTT-3') and D3B (5'-TCGGAAGGAACCAGCTACTA-3') (1). PCR products were purified and sequenced at the Macrogene sequencing facility service (Amsterdam, The Netherlands). Sequences were deposited in GenBank (KJ645427 to 33). The sequences were compared with previously published sequences in NCBI database and showed 96 to 100% sequence similarity with M. hapla accession nos. GQ130139, DQ328685, KF430798, and DQ145641. Unfortunately, comparison of sequences did not provide further information about the origin of this Ethiopian population, if it is native to Ethiopia or was imported with infected plant material. To the best of our knowledge, this is the first record of M. hapla occurring in Ethiopia. M. hapla is known as a serious pest of roses in colder climate regions. In Africa, it was previously reported from Tanzania (3) and South Africa (2). Thus, it appears that this species has now become also established in Ethiopia at higher altitudes (1,400 to 2,100 m above sea level) within the urban hinterland of Addis Ababa. References: (1) Baldwin et al. Mol. Phy. Evol. 8:248, 1887. (2) J. H. O'Bannon. Institute Agri. Res. 29, 1975. (3) E. Onkendi and L. N. Moleleki. Eur. J. Pl. Pathol. 136:1, 2013. (4) A. G. Whitehead. Trans. Zool. Soc. Lon.31:263, 1968.

Single pulse coherence measurements in the water window at the free-electron laser FLASH.

The spatial coherence of free-electron laser radiation in the water window spectral range was studied, using the third harmonic (λ<(3rd) = 2.66 nm) of DESY's Free-electron LASer in Hamburg (FLASH). Coherent single pulse diffraction patterns of 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) multilamellar lipid stacks have been recorded. The intensity histogram of the speckle pattern around the first lamellar Bragg peak, corresponding to the d = 5 nm periodicity of the stack, reveals an average number of transverse modes of M¯ = 3.0 of the 3rd harmonic. Using the lipid stack as a 'monochromator', pulse-to-pulse fluctuations in the third harmonic λ(3rd) have been determined to be 0.033 nm.

Pump-probe X-ray holographic imaging of laser-induced cavitation bubbles with femtosecond FEL pulses.

Cavitation bubbles can be seeded from a plasma following optical breakdown, by focusing an intense laser in water. The fast dynamics are associated with extreme states of gas and liquid, especially in the nascent state. This offers a unique setting to probe water and water vapor far-from equilibrium. However, current optical techniques cannot quantify these early states due to contrast and resolution limitations. X-ray holography with single X-ray free-electron laser pulses has now enabled a quasi-instantaneous high resolution structural probe with contrast proportional to the electron density of the object. In this work, we demonstrate cone-beam holographic flash imaging of laser-induced cavitation bubbles in water with nanofocused X-ray free-electron laser pulses. We quantify the spatial and temporal pressure distribution of the shockwave surrounding the expanding cavitation bubble at time delays shortly after seeding and compare the results to numerical simulations.

Diffraction properties of periodic lattices under free electron laser radiation.

In this Letter, we report the pioneering use of free electron laser radiation for the investigation of periodic crystalline structures. The diffraction properties of silver behenate single nanocrystals (5.8 nm periodicity) with the dimensions of 20 nm x 20 nm x 20 microm and as powder with grain sizes smaller than 200 nm were investigated with 8 nm free electron laser radiation in single-shot modus with 30 fs long free electron laser pulses. This work emphasizes the possibility of using soft x-ray free electron laser radiation for these crystallographic studies on a nanometer scale.

Development of a hard X-ray split-and-delay line and performance simulations for two-color pump-probe experiments at the European XFEL.

A hard X-ray Split-and-Delay Line (SDL) under construction for the Materials Imaging and Dynamics station at the European X-Ray Free-Electron Laser (XFEL) is presented. This device aims at providing pairs of X-ray pulses with a variable time delay ranging from -10 ps to 800 ps in a photon energy range from 5 to 10 keV for photon correlation and X-ray pump-probe experiments. A custom designed mechanical motion system including active feedback control ensures that the high demands for stability and accuracy can be met and the design goals achieved. Using special radiation configurations of the European XFEL's SASE-2 undulator (SASE: Self-Amplified Spontaneous Emission), two-color hard x-ray pump-probe schemes with varying photon energy separations have been proposed. Simulations indicate that more than 109 photons on the sample per pulse-pair and up to about 10% photon energy separation can be achieved in the hard X-ray region using the SDL.

Endophytic Colonization of Plants by the Biocontrol Agent Rhizobium etli G12 in Relation to Meloidogyne incognita Infection.

ABSTRACT The external and internal colonization of potato and Arabidopsis roots by the biocontrol strain Rhizobium etli G12 containing a plasmidborne trp promoter green fluorescent protein transcriptional fusion, pGT-trp, was studied in the presence and absence of the root-knot nematode Meloidogyne incognita. Plant colonization behavior and biocontrol potential of the marked strain G12(pGT-trp) was not altered compared with the parental strain. Plasmid pGT-trp was stable for more than 80 generations without selection and conferred sufficient fluorescence to detect single bacterial cells in planta. Although bacteria were found over the entire rhizoplane, they preferentially colonized root tips, the emerging lateral roots, and galled tissue caused by Meloidogyne infestation. Internal colonization of potato roots was mainly observed in epidermal cells, especially root hairs. G12(pGT-trp) colonization was also observed in inner Arabidopsis root tissues in areas of vascularization. In the presence of M. incognita, G12(pGT-trp) colonized the interior of nematode galls in high numbers. In some cases, bacterial colonization even extended from the galled tissue into adjacent root tissue. The internally colonized sites in roots were often discontinuous. Fluorescence microscopy of gfp-tagged rhizobacteria was a sensitive and a rapid technique to study external and internal colonization of plant roots by bacteria interacting with nematodes.

Activity and Differential Induction of Chitinase Isozymes in Soybean Cultivars Resistant or Susceptible to Root-knot Nematodes.

Host physiological events in relation to infestation by parasitic nematodes are not well documented. Soybean plant responses to Meloidogyne incognita infestation were compared to resistant (Bryan) and susceptible (Brim) cultivars at 0, 1, 3, 10, 20, and 34 days after infestation (DAI). The resistant cultivar had higher chitinase activity than the susceptible cultivar at every sample time beginning at 3 DAI. Results from isoelectric focusing gel electrophoresis analyses indicated that three acidic chitinase isozymes with isoelectric points (pIs) of 4.8, 4.4, and 4.2 accumulated to a greater extent in the resistant compared to the susceptible cultivar following challenge. SDS-PAGE analysis of root proteins revealed that two proteins with molecular weights of approximately 31 and 46 kD accumulated more rapidly and to a higher level in the resistant than in the susceptible cultivar. Additionally, three major protein bands (33, 22, and 20 kD) with chitinase activity were detected with a modified SDS-PAGE analysis in which glycolchitin was added into the gel matrix. These results indicate that higher chitinase activity and early induction of specific chitinase isozymes may be associated with resistance to root-knot nematode in soybean.

Development of a multipurpose vacuum chamber for serial optical and diffraction experiments with free electron laser radiation.

In this paper we present a development of a multipurpose vacuum chamber which primal function is to be used in pump/probe experiments with free electron laser (FEL) radiation. The chamber is constructed for serial diffraction and serial spectroscopy allowing a fast exchange of samples during the measurement process. For the fast exchange of samples, liquid jet systems are used. Both applications, utilizing soft x-ray FEL pulses as pump and optical laser pulses as probe and vice versa are documented. Experiments with solid samples as well as the liquid jet samples are presented. When working with liquid jets, a system of automatically refilled liquid traps for capturing liquids has been developed in order to ensure stable vacuum conditions. Differential pumping stages are placed in between the FEL beamline and the experimental chamber so that working pressure in the chamber can be up to four orders of magnitude higher than the pressure in the FEL beamline.

Influence of plant species on the biological control activity of the antagonistic rhizobacterium Rhizobium etli strain G12 toward the root-knot nematode, Meloidogyne incognita.

The influence of plant species on the antagonistic activity of the rhizosphere bacterium Rhizobium etli G12 towards the root-knot nematode Meloidogyne incognita was studied. The crops tested were tomato, cucumber, cotton, soybean and pepper. The plants were evaluated for the following parameters: root gall-index, total number of galls and egg masses of M. incognita, as well as shoot and root fresh weight and root length. Results indicated a clear influence of plant species on the ability of R. etli G12 to reduce nematode infection. Based on the root gall index, nematode control by R. etli G12 was higher on vegetables (tomato, cucumber, pepper) than on field crops (soybean, cotton). Reduction in galling ranged from 17% for cotton to 50% for tomato. R. etli G12 also reduced the actual number of galls produced. The reduction in the number of galls produced between crops was not affected significantly as was seen when a galling index was used to measure activity. The reduction in the number of galls was similar in level for all the crops studied and ranged from 34% for cucumber to 47% for tomato. There was a significant reduction in the number of egg masses produced by the females ranging from 37% for soybean to 70% for pepper. This indicated a direct effect on female development in the root after penetration or delayed penetration on certain crops. The bacteria caused significant increases in shoot fresh weight from 11% for soybean to 31% for pepper and in root fresh weight from 3% for soybean to 39% for tomato and in root length from 11% for cucumber to 24% for pepper. R. etli G12 gave significant control of M. incognita on a broad range of host plants, but the level of control varied. The suitability of each plant species, therefore, must be examined before R. etli G12 can be recommend for control of this nematode.

Induced systemic resistance of selected endophytic bacteria against Meloidogyne incognita on tomato.

In previous work, the four endophytic bacteria Pantoea agglomerans MK-29, Cedeca davisae MK-30, Enterobacter spp. MK-42 and Pseudomonas putida MT-19 were shown to reduce Meloidogyne incognita on tomato when applied as a seed treatment and/or soil drench. The objective of this work was to study these bacteria for their potential to induce systemic resistance against root knot nematodes on tomato. To guarantee spatial separation between inducing agent and pathogen a split-root system was chosen and inoculated with the bacteria as a drench application on one side of the root system and 6 days later with 2000 juveniles of Meloidogyne incognita on the other side of the split-root system. The experiment was maintained in the greenhouse and repeated once. The penetration rate of juveniles as well as the total number of root-knot galls and egg masses was recorded. Treatment with all four bacteria significantly reduced juvenile penetration and the number of root-knot galls when compared with the non-treated control. Induced systemic resistance is considered a possible control mechanism of endophytic bacteria against root-knot nematodes.

Effects of natural products on soil organisms and plant health enhancement.

TerraPy, Magic Wet and Chitosan are soil and plant revitalizers based on natural renewable raw materials. These products stimulate microbial activity in the soil and promote plant growth. Their importance to practical agriculture can be seen in their ability to improve soil health, especially where intensive cultivation has shifted the biological balance in the soil ecosystem to high numbers of plant pathogens. The objective of this study was to investigate the plant beneficial capacities of TerraPy, Magic Wet and Chitosan and to evaluate their effect on bacterial and nematode communities in soils. Tomato seedlings (Lycopersicum esculentum cv. Hellfrucht Frühstamm) were planted into pots containing a sand/soil mixture (1:1, v/v) and were treated with TerraPy, Magic Wet and Chitosan at 200 kg/ha. At 0, 1, 3, 7 and 14 days after inoculation the following soil parameters were evaluated: soil pH, bacterial and fungal population density (cfu/g soil), total number of saprophytic and plant-parasitic nematodes. At the final sampling date tomato shoot and root fresh weight as well as Meloidogyne infestation was recorded. Plant growth was lowest and nematode infestation was highest in the control. Soil bacterial population densities increased within 24 hours after treatment between 4-fold (Magic Wet) and 19-fold (Chitosan). Bacterial richness and diversity were not significantly altered. Dominant bacterial genera were Acinetobacter (41%) and Pseudomonas (22%) for TerraPy, Pseudomonas (30%) and Acinetobacter (13%) for Magic Wet, Acinetobacter (8.9%) and Pseuodomonas (81%) for Chitosan and Bacillus (42%) and Pseudomonas (32%) for the control. Increased microbial activity also was associated with higher numbers of saprophytic nematodes. The results demonstrated the positive effects of natural products in stimulating soil microbial activity and thereby the antagonistic potential in soils leading to a reduction in nematode infestation and improved plant growth.

Lipopolysaccharides of Rhizobium etli strain G12 act in potato roots as an inducing agent of systemic resistance to infection by the cyst nematode Globodera pallida.

Recent studies have shown that living and heat-killed cells of the rhizobacterium Rhizobium etli strain G12 induce in potato roots systemic resistance to infection by the potato cyst nematode Globodera pallida. To better understand the mechanisms of induced resistance, we focused on identifying the inducing agent. Since heat-stable bacterial surface carbohydrates such as exopolysaccharides (EPS) and lipopolysaccharides (LPS) are essential for recognition in the symbiotic interaction between Rhizobium and legumes, their role in the R. etli-potato interaction was studied. EPS and LPS were extracted from bacterial cultures, applied to potato roots, and tested for activity as an inducer of plant resistance to the plant-parasitic nematode. Whereas EPS did not affect G. pallida infection, LPS reduced nematode infection significantly in concentrations as low as 1 and 0.1 mg ml(-1). Split-root experiments, guaranteeing a spatial separation of inducing agent and challenging pathogen, showed that soil treatments of one half of the root system with LPS resulted in a highly significant (up to 37%) systemic induced reduction of G. pallida infection of potato roots in the other half. The results clearly showed that LPS of R. etli G12 act as the inducing agent of systemic resistance in potato roots.

Effects of arbuscular mycorrhizal fungi and a non-pathogenic Fusarium oxysporum on Meloidogyne incognita infestation of tomato.

Arbuscular mycorrhizal (AM) fungi and non-pathogenic strains of soil-borne pathogens have been shown to control plant parasitic nematodes. As AM fungi and non-pathogenic fungi improve plant health by different mechanisms, combination of two such partners with complementary mechanisms might increase overall control efficacy and, therefore, provide an environmentally safe alternative to nematicide application. Experiments were conducted to study possible interactions between the AM fungus Glomus coronatum and the non-pathogenic Fusarium oxysporum strain Fo162 in the control of Meloidogyne incognita on tomato. Pre-inoculation of tomato plants with G. coronatum or Fo162 stimulated plant growth and reduced M. incognita infestation. Combined application of the AM fungus and Fo162 enhanced mycorrhization of tomato roots but did not increase overall nematode control or plant growth. A higher number of nematodes per gall was found for mycorrhizal than non-mycorrhizal plants. In synergisms between biocontrol agents, differences in their antagonistic mechanisms seem to be less important than their effects on different growth stages of the pathogen.