Effect of climate change on sporulation of the teleomorphs of <Emphasis Type="Italic">Leptosphaeria</Emphasis> species causing stem canker of brassicas

Leptosphaeria maculans and L. biglobosa are closely related sibling fungal pathogens that cause phoma leaf spotting, stem canker (blackleg) and stem necrosis of oilseed rape (Brassica napus). The disease is distributed worldwide, and it is one of the main causes of considerable decrease in seed yield and quality. Information about the time of ascospore release at a particular location provides important data for decision making in plant protection, thereby enabling fungicides to be used only when necessary and at optimal times and doses. Although the pathogens have been studied very extensively, the effect of climate change on the frequencies and distributions of their aerially dispersed primary inoculum has not been reported to date. We have collected a large dataset of spore counts from Poznan, located in central-west part of Poland, and studied the relationships between climate and the daily concentrations of airborne propagules over a period of 17 years (1998–2014). The average air temperature and precipitation for the time of development of pseudothecia and ascospore release (July–November), increased during the years under study at the rates of 0.1 °C and 6.3 mm per year. The day of the year (DOY) for the first detection of spores, as well as the date with maximum of spores, shifted from 270 to 248 DOY, and from 315 to 265 DOY, respectively. The acceleration of the former parameter by 22 days and the latter by 50 days has great influence on the severity of stem canker of oilseed rape.     

Further evidence for the absence of polyproline II stretch in the XAO peptide

It has been suggested that the alanine-based peptide with sequence Ac-XX-[A](7)-OO-NH(2), termed XAO where X denotes diaminobutyric acid and O denotes ornithine, exists in a predominantly polyproline-helix (P(II)) conformation in aqueous solution. In our recent work, we demonstrated that this "polyproline conformation" should be regarded as a set of local conformational states rather than as the overall conformation of the molecule. In this work, we present further evidence to support this statement. Differential scanning calorimetry measurements showed only a very small peak in the heat capacity of an aqueous solution of XAO at 57 degrees C, whereas the suggested transition to the P(II) structure should occur at approximately 30 degrees C. We also demonstrate that the temperature dependence of the (3)J(HNHalpha) coupling constants of the alanine residues can be explained qualitatively in terms of Boltzmann averaging over all local conformational states; therefore, this temperature dependence proves that a conformational transition does not occur. Canonical MD simulations with the solvent represented by the generalized Born model, and with time-averaged NMR-derived restraints, demonstrate the presence of an ensemble of structures with a substantial amount of local P(II) conformational states but not with an overall P(II) conformation.     

Kinetics and Equilibrium Studies of Tet Repressor–Operator Interaction

Binding of a Tet repressor mutant containing a single Trp43 residue in the tet operator recognition α-helix leads to the quenching of the protein fluorescence down to about 23% in the case of the tet O₁ operator and to 40% in the case of the tet O₂ operator. We have used fluorescence detection to describe the binding equilibrium and kinetics of the Tet repressor interaction with the 20-bp DNA operators tet O₁ and tet O₂. Stopped-flow measurements in an excess of the tet operators performed in 5 mM NaCl or 150 mM NaCl indicate that the reaction can be described by at least three exponentials characterized by different relaxation times. The mechanism of interaction for both operators as well as for two salt concentrations used can be described as TetR + Operator ? Complex 1 ? Complex 2 ? Complex 3. Only the much faster process can be described as a second-order reaction characterized by a bimolecular rate constant equal to 2.8 × 10⁶ M^?1 sec^?1 for both operators. The medium and slow processes may be described by relaxational times ranging from 50 msec to seconds. The results of the binding equilibrium measurements extrapolated to 1 M NaCl concentration, which reflects the specific nonionic interaction between TetR and tet operators, indicate K _as equal to 3.2 × 10⁴ and 4.0 × 10⁵ M^?1 for tet O₁ and tet O₂, respectively. The number of monovalent ions replaced upon binding can be calculated as about 5 and 3 for tet O₁ and tet O₂, respectively. The binding of Tet repressor to the operators leads to changes in the circular dichroism spectra of the DNA which could indicate transitions of B-DNA into A-like DNA structure.     

Structural studies of the C‐terminal 19‐peptide of serum amyloid A and its Pro→Ala variants interacting with human cystatin C

Serum amyloid A (SAA) is a multifunctional acute‐phase protein whose concentration in serum increases markedly following a number of chronic inflammatory and neoplastic diseases. Prolonged high SAA level may give rise to reactive systemic amyloid A (AA) amyloidosis, where the N‐terminal segment of SAA is deposited as amyloid fibrils. Besides, recently, well‐documented association of SAA with high‐density lipoprotein or glycosaminoglycans, in particular heparin/heparin sulfate (HS), and specific interaction between SAA and human cystatin C (hCC), the ubiquitous inhibitor of cysteine proteases, was proved. Using a combination of selective proteolytic excision and high‐resolution mass spectrometry, a hCC binding site in the SAA sequence was determined as SAA(86–104). The role of this SAA C‐terminal fragment as a ligand‐binding locus is still not clear. It was postulated important in native SAA folding and in pathogenesis of AA amyloidosis. In the search of conformational details of this SAA fragment, we did its structure and affinity studies, including its selected double/triple Pro→Ala variants. Our results clearly show that the SAA(86–104) 19‐peptide has rather unordered structure with bends in its C‐terminal part, which is consistent with the previous results relating to the whole protein. The results of affinity chromatography, fluorescent ELISA‐like test, CD and NMR studies point to an importance of proline residues on structure of SAA(86–104). Conformational details of SAA fragment, responsible for hCC binding, may help to understand the objective of hCC–SAA complex formation and its importance for pathogenesis of reactive amyloid A amyloidosis. Copyright © 2015 John Wiley & Sons, Ltd.     

Morphological and molecular examination of relationships and epitype establishment of Phacus pleuronectes,Phacus orbicularis,and Phacus hamelii1

Verification of morphological diagnostic features and the establishment of three epitypes for three species of Phacus Dujardin—Phacus pleuronectes (O. F. Müll.) Dujardin, Phacus orbicularis Hübner, and Phacus hamelii Allorge et Lefèvre—was performed based on literature studies and analysis of morphological (cell shape, cell size, and periplast ornamentation) as well as molecular (18S rDNA) characters. Periplast ornamentation was recognized as a main diagnostic character, distinguishing P. orbicularis from P. pleuronectes and P. hamelii. Phacus orbicularis has struts running perpendicular to the longitudinal axis of the strips, while P. pleuronectes and P. hamelii do not. On the SSU rDNA tree, obtained by the Bayesian method, P. orbicularis, P. pleuronectes, and P. hamelii belong to three distinct clades. Some of the phylogenetic relationships are not resolved, but there are at least three Phacus species (P. hamatus, P. platyaulax, P. longicauda; for taxonomic authors, see Introduction) that are more closely related to P. orbicularis than is P. pleuronectes. Phacus hamelii is more closely related to P. ranula and the assemblage of several species of Phacus, which have small cells, than to P. orbicularis or P. pleuronectes.