The vacuolar chitinases of class I possess an N-terminal cysteine-rich domain homologous to hevein and chitin-binding lectins such as wheat germ agglutinin and Urtica dioica lectin. To investigate the significance of this domain for the biochemical and functional characteristics of chitinase, chimeric genes encoding the basic chitinase A of tobacco (Nicotiana tabacum) with and without this domain were constructed and constitutively expressed in transgenic Nicotiana sylvestris. The chitinases were subsequently isolated and purified to homogeneity from the transgenic plants. Chromatography on colloidal chitin revealed that only the form with the N-terminal domain, and not the one without it, had chitin-binding properties, demonstrating directly that the domain is a chitin-binding domain (CBD). Under standard assay conditions with radioactive colloidal chitin, both forms of chitinase had approximately the same catalytic activity. However, kinetic analysis demonstrated that the enzyme without CBD had a considerably lower apparent affinity for its substrate. The pH and temperature optima of the two chitinases were similar, but the form with the CBD had an approximately 3-fold higher activation energy and retained a higher activity at low pH values. Both chitinases were capable of inhibiting growth of Trichoderma viride, although the form with the CBD was about three times more effective than the one without it. Thus, the CBD is not necessary for catalytic or antifungal activity of chitinase.
A PCR assay which allows detection and quantification of Epichloë endophytes in tissues of the grass Bromus erectus is described. PCR with specific primers flanking a microsatellite-containing locus (MS primers) amplified fragments 300 to 400 bp in length from as little as 1.0 pg of fungal genomic DNA in 100 ng of DNA from infected plant material. When annealing temperatures were optimized, all Epichloë and Acremonium strains tested, representing many of the known taxonomic groups, yielded an amplification product, indicating that the MS primers may be useful for in planta detection of a variety of related species, including agronomically important Acremonium coenophialum and Acremonium lolii. No fragments were generated from DNA isolates from uninfected plant material or from unrelated fungi isolated from B. erectus. For diagnostic applications, a B. erectus-specific primer pair was designed for use in multiplex PCR to allow simultaneous amplification of plant and fungal DNA sequences, providing an internal control for PCR failure caused by inhibitory plant compounds present in DNA extracts. For quantitative applications, a heterologous control template in primer binding sites complementary to the MS primers was constructed for use in competitive PCR, allowing direct quantification of Epichloë in plant DNA extracts. The fungal DNA present in infected leaves of B. erectus between 1 and 20 pg per 100 ng of leaf DNA, but the amounts of fungal DNA present in the sheath and blade of a given leaf were correlated, indicating that the degree of infection varied between plant individuals but that leaves were colonized in a uniform way.
During its calibration and performance verification phase, the eROSITA instrument aboard the Spectrum-RG satellite performed a uniform wide-area X-ray survey of approximately 140 deg2, known as the eROSITA Final Equatorial Depth Survey (eFEDS). The primary aim of eFEDS is to demonstrate the scientific performance to be expected at the end of the eight--pass eROSITA all--sky survey. This survey will provide the first focussed image of the whole sky in the hard X-ray ($>2$ keV) bandpass. The expected source population in this energy range is thus of great interest, particularly for AGN studies. We used a 2.3--5 keV selection to construct a sample of 246 point-like hard X-ray sources for further study and characterisation. These sources are classified as either extragalactic ($ 90$ <!PCT!>) or Galactic ($ 10$ <!PCT!>), with the former consisting overwhelmingly of AGN and the latter active stars. We concentrated our further analysis on the extragalactic AGN sample, describing their X-ray and multi-wavelength properties and comparing them to the eFEDS main AGN sample selected in the softer 0.2--2.3 keV band. The eROSITA hard band selects a subsample of sources that is a factor of more than ten brighter than the eFEDS main sample. The AGN within the hard population reach up to $z=3.2$ but on the whole, they are relatively nearby, with median $z$=0.34 compared to $z$=0.94 for the main sample. The hard survey probes typical luminosities in the range $ X = 43-46$. The X-ray spectral analysis shows significant intrinsic absorption (with $ H >21$) in $ 20$ <!PCT!> of the sources, with a hard X-ray power law continuum with mean $< which is typical of AGN, but slightly harder than the soft-selected eROSITA sample. Around $10$ <!PCT!> of the hard sample show a significant `soft excess' component. The sampled black hole mass distribution in the eFEDS broad-line AGN population is consistent with that of the deeper COSMOS survey that probes a higher redshift population. On the other hand, the Eddington ratios appear systematically lower, which is consistent with the idea that the decline in SMBH activity since $z 1$ is due to a reduction in the typical accretion rate, rather than a shift towards activity in lower-mass black holes. The eFEDS hard sample provides a preview of what can be expected from the eRASS final survey in terms of data quality. This pilot survey indicates the power of eROSITA to shed new light on the demographics and evolution of AGN, and the potential for discovery of new and rare populations.
We present a spectral analysis of the narrow-line Seyfert 1 galaxy I Zwicky 1, focusing on the characteristics of the ionized absorbers as observed with XMM–Newton in 2005. The soft X-ray spectrum shows absorption by two components of ionized gas with a similar column density (NH∼ 1021 cm−2) and ionization parameters log ξ∼ 0 and 2.5. Comparing this observation with a 2002 XMM–Newton data set, we see a clear anticorrelation between the X-ray ionization parameter ξX and the 0.1–10 keV luminosity. Viable explanations for this effect include transient clouds or filaments crossing the line of sight in a complex geometry or a gas observed in non-equilibrium. The outflow velocity of the X-ray low-ionization absorber is consistent with the outflow of the ultraviolet (UV) absorber detected in a past Hubble Space Telescope observation. In addition, the ionic column densities of C iv and N v derived from the X-ray model are consistent with the UV values. This suggests that the low-ionization outflowing gas may survive for many years, despite large changes in flux, and that there is a tight connection between the X-ray and UV absorbers that can only be confirmed with a simultaneous UV and X-ray observation.
lant scientists are easy to identify, one might think. That’s surely true when comparing them to the sometimes rath-er colourful variety of people gathered under the umbrellas of certain other biodisciplines. Okay, plant scientists do not only work in explicit plant science institutes; you’ll find them in bio-chemistry, genetics and developmental biology institutes as well, although that doesn’t matter. Actually, you only have to check which organism each researcher uses for her or his experiments – and you’ll easily filter out the plant scientists. Well, that certainly makes sense. Nevertheless, you might still encounter a problem or two. What to do for example with the structural biologist who particularly likes to pick photosynthet-ic membrane proteins for his NMR studies? Or the computation-al biologist who has just co-published a couple of software tools designed for the analysis of plant genome sequences? Are they “plant scientists”?We said ‘no’ for two main reasons. Firstly, we think that those people don’t place their main research focus on
Based on previous publications exploring pseudo-complex General Relativity (pc-GR) we present a selection of observable consequences of pc-GR and possible ways to experimentally access them. Whenever possible we compare the results to Einstein's GR and differences are worked out in detail. We propose experimental tests to check the predictions of pc-GR for the orbital frequency of test particles, the gravitational redshift effect and the last stable orbit. We will show that the orbital frequency of test particles at a given radius in pc-GR is in general lower compared to standard GR. Also the effect of frame dragging is modified (weakened) in pc-GR. Concerning the gravitational redshift of a radiation emitting object we find that it is also lower in pc-GR than in standard GR. Eventually the classical concept of a last stable orbit has to be modified in pc-GR.
'%3e%3cpath fill='%23012169' d='M0 0v30h60V0z'/%3e%3cpath stroke='%23fff' stroke-width='6' d='m0 0 60 30m0-30L0 30'/%3e%3cpath stroke='%23C8102E' stroke-width='4' d='m0 0 60 30m0-30L0 30' clip-path='url(%23b)'/%3e%3cpath stroke='%23fff' stroke-width='10' d='M30 0v30M0 15h60'/%3e%3cpath stroke='%23C8102E' stroke-width='6' d='M30 0v30M0 15h60'/%3e%3c/g%3e%3c/svg%3e)