<Emphasis Type="Italic">Sphaerophragmium pulchrum</Emphasis>, a new species of rust fungi on <Emphasis Type="Italic">Albizia adinocephala</Emphasis> from Panama

Sphaerophragmium pulchrum is proposed as a new microcyclic rust species on Albizia adinocephala from Panama. Telia and spermogonia are described, illustrated, and compared to those of known species of Sphaerophragmium. Spermogonia are described for the first time for a species of Sphaerophragmium. This is the first record of this genus for the southern part of the Central American isthmus.     

Novel Type of ADP-Forming Acetyl Coenzyme A Synthetase in Hyperthermophilic Archaea: Heterologous Expression and Characterization of Isoenzymes from the Sulfate Reducer Archaeoglobus fulgidus and the Methanogen Methanococcus jannaschii

Acetyl coenzyme A (CoA) synthetase (ADP forming) (ACD) represents a novel enzyme of acetate formation and energy conservation (acetyl-CoA + ADP + P(i) right harpoon over left harpoon acetate + ATP + CoA) in Archaea and eukaryotic protists. The only characterized ACD in archaea, two isoenzymes from the hyperthermophile Pyrococcus furiosus, constitute 145-kDa heterotetramers (alpha(2), beta(2)). The coding genes for the alpha and beta subunits are located at different sites in the P. furiosus chromosome. Based on significant sequence similarity of the P. furiosus genes, five open reading frames (ORFs) encoding putative ACD were identified in the genome of the hyperthermophilic sulfate-reducing archaeon Archaeoglobus fulgidus and one ORF was identified in the hyperthermophilic methanogen Methanococcus jannaschii. The ORFs constitute fusions of the homologous P. furiosus genes encoding the alpha and beta subunits. Two ORFs, AF1211 and AF1938, of A. fulgidus and ORF MJ0590 of M. jannaschii were cloned and functionally overexpressed in Escherichia coli. The purified recombinant proteins were characterized as distinctive isoenzymes of ACD with different substrate specificities. In contrast to the Pyrococcus ACD, the ACDs of Archaeoglobus and Methanococcus constitute homodimers of about 140 kDa composed of two identical 70-kDa subunits, which represent fusions of the homologous P. furiosus alpha and beta subunits in an alphabeta (AF1211 and MJ0590) or betaalpha (AF1938) orientation. The data indicate that A. fulgidus and M. jannaschii contains a novel type of ADP-forming acetyl-CoA synthetase in Archaea, in which the subunit polypeptides and their coding genes are fused.     

New species and new records of smut fungi from China

This paper deals with six species of smut fungi which were collected in the provinces Yunnan and Gansu in China. Among them, Anthracoidea yunnanensis and Entorrhiza guttiformis are new species. Sporisorium andropogonis-annulati, which is a new combination, Sporisorium montaniensis, Urocystis pseudoanemones, and Ustilago griffithsii are new records for China.     

Molecular phylogeny and host-plant use (Lamiaceae) of the Thymogethes pollen beetles (Coleoptera)

The 24 members of the Euro-Asiatic genus Thymogethes are highly specialized pollen beetles associated as larvae with flowers of Lamiaceae Nepetoideae. All members of the genus were analysed in within the framework of an integrative taxonomy approach, which was aimed to reconstruct the phylogenetic relationships, as well as the possible pattern of evolution of their larval-host-plant association. Evidence from multiple molecular markers [COI; 16S; H3], combined with an estimation of divergence times using an average rate of 0.0177 substitutions/site/My among branches, placed the origin of the genus at a minimum of 9–10 Mya. This date of origin approximates the known evolution of the host plants in Euro-Mediterranean areas. Evidence from combined molecular and cladistic morphological analyses resulted in suitable agreement with the previously established morphology-based systematics of the genus, although members of the exilis species-group were split into three clades. The only disagreement between results of this new combined phylogeny and previous classification is in the exclusion of “Thymogethes” grenieri. This species is herein positioned outside the genus, based on molecular evidence. Our analysis depicts several Thymogethes species differentiating in the last few Mys, specifically those included in the T. lugubris species-group. Combined evidence from DNA, morphology and ancestral state parsimony reconstruction of larval-host-plant associations suggests that subtribe Menthinae likely represents the ancestral host plants, with a series of independent host shifts during the radiation of the clade, in association first with Menthinae and subsequently with Lavandulinae and Nepetinae. Steno-oligophagy is the most frequent (86%) condition, while strictly monophagous species are less numerous (14%).     

Molecular modelling of the mass density of single proteins

Using molecular dynamics (MD) simulations, the density of single proteins and its temperature dependence was modelled starting from the experimentally determined protein structure and a generic, transferable force field, without the need of prior parameterization. Although all proteins consist of the same 20 amino acids, their density in aqueous solution varies up to 10% and the thermal expansion coefficient up to twofold. To model the protein density, systematic MD simulations were carried out for 10 proteins with a broad range of densities (1.32–1.43?g/cm³) and molecular weights (7–97?kDa). The simulated densities deviated by less than 1.4% from their experimental values that were available for four proteins. Further analyses of protein density showed that it can be essentially described as a consequence of amino acid composition. For five proteins, the density was simulated at different temperatures. The simulated thermal expansion coefficients ranged between 4.3 and 7.1?×?10^?4?K^?1 and were similar to the experimentally determined values of ribonuclease-A and lysozyme (deviations of 2.4 and 14.6%, respectively). Further analyses indicated that the thermal expansion coefficient is linked to the temperature dependence of atomic fluctuations: proteins with a high thermal expansion coefficient show a low increase in flexibility at increasing temperature. A low increase in atomic fluctuations with temperature has been previously described as a possible mechanism of thermostability. Thus, a high thermal expansion coefficient might contribute to protein thermostability.     

Membrane Region M2C2 in Subunit KtrB of the K+ Uptake System KtrAB from Vibrio alginolyticus Forms a Flexible Gate Controlling K+ Flux: AN ELECTRON PARAMAGNETIC RESONANCE STUDY*

Transmembrane stretch M_2C from the bacterial K⁺-translocating protein KtrB is unusually long. In its middle part, termed M_2C2, it contains several small and polar amino acids. This region is flanked by the two α-helices M_2C1 and M_2C3 and may form a flexible gate at the cytoplasmic side of the membrane controlling K⁺ translocation. In this study, we provide experimental evidence for this notion by using continuous wave and pulse EPR measurements of single and double spin-labeled cysteine variants of KtrB. Most of the spin-labeled residues in M_2C2 were shown to be immobile, pointing to a compact structure. However, the high polarity revealed for the microenvironment of residue positions 317, 318, and 327 indicated the existence of a water-accessible cavity. Upon the addition of K⁺ ions, M_2C2 residue Thr-318R1 (R1 indicates the bound spin label) moved with respect to M_2B residue Asp-222R1 and M_2C3 residue Val-331R1 but not with respect to M_2C1 residue Met-311R1. Based on distances determined between spin-labeled residues of double-labeled variants of KtrB in the presence and absence of K⁺ ions, structural models of the open and closed conformations were developed.     

Molecular mechanisms underpinning phosphorus‐use efficiency in rice

Orthophosphate (H₂PO₄^?, Pi) is an essential macronutrient integral to energy metabolism as well as a component of membrane lipids, nucleic acids, including ribosomal RNA, and therefore essential for protein synthesis. The Pi concentration in the solution of most soils worldwide is usually far too low for maximum growth of crops, including rice. This has prompted the massive use of inefficient, polluting, and nonrenewable phosphorus (P) fertilizers in agriculture. We urgently need alternative and more sustainable approaches to decrease agriculture's dependence on Pi fertilizers. These include manipulating crops by (a) enhancing the ability of their roots to acquire limiting Pi from the soil (i.e. increased P‐acquisition efficiency) and/or (b) increasing the total biomass/yield produced per molecule of Pi acquired from the soil (i.e. increased P‐use efficiency). Improved P‐use efficiency may be achieved by producing high‐yielding plants with lower P concentrations or by improving the remobilization of acquired P within the plant so as to maximize growth and biomass allocation to developing organs. Membrane lipid remodelling coupled with hydrolysis of RNA and smaller P‐esters in senescing organs fuels P remobilization in rice, the world's most important cereal crop.     

Effects of insecticide application on parasitism rates of pollen beetle larvae (<Emphasis Type="Italic">Brassicogethes aeneus</Emphasis> (Fabricius)) by tersilochine parasitoids

Larval parasitoids can substantially reduce the population density of the pollen beetle [Brassicogethes aeneus (Fabricius), syn. Meligethes aeneus (Fabricius)]. The most abundant tersilochine parasitoids of pollen beetle are Tersilochus heterocerus, Phradis interstitialis and P. morionellus. The main activity of these parasitoids was observed in the period shortly before flowering to full flowering of oilseed rape. Insecticide applications during this period may have negative effects on parasitoids. In the present study, the effects of the insecticides Biscaya (a.i. thiacloprid), Mavrik (a.i. tau-fluvalinate) and Karate Zeon (a.i. lambda-cyhalothrin) applied during the bud or flowering stage of winter oilseed rape on parasitization of pollen beetle larvae by T. heterocerus were studied in 12 field trials at different locations in Germany in 2013–2015. The effects on parasitism by Phradis spp. were assessed in 2015. Parasitism of pollen beetle larvae by T. heterocerus was found in all field trials in all experimental years, but in most trials not before full flowering. Maximum percentage of parasitized larvae at different locations ranged between 3.4 and 16.8% in 2013, 8.3 and 22.4% in 2014 and from 11.1 to 29.1% in 2015. Levels of parasitism were not significantly different between the untreated control and insecticide treatments within each location. In contrast to T. heterocerus, Phradis spp. was not detected at all locations and not before flowering declining. In field trials at Lucklum and Puch, the maximum level of parasitism by Phradis spp. was 9.4 and 18.3%, respectively. No significant effect of insecticide application on parasitism by Phradis spp. was observed between the treatments. The results of this study showed that the insecticides used in the field trials did not affect parasitization of pollen beetle larvae by T. heterocerus and Phradis spp., regardless whether applied at the bud stage, at the beginning of flowering or full flowering.     

Impact of insecticides on oilseed rape bud infestation with eggs and larvae of pollen beetle (<Emphasis Type="Italic">Brassicogethes aeneus</Emphasis> (Fabricius))

One of the main insect pests in oilseed rape is the pollen beetle (Brassicogethes aeneus (Fabricius), syn. Meligethes aeneus). To maximize efficiency of control of this pest, insecticides are required that ideally, not just prevent yield losses by bud feeding of overwintered pollen beetles, but simultaneously minimize the reproduction of the pest, thereby reducing the size of the following generation infesting next year’s oilseed rape. The neonicotinoid active substance thiacloprid is known to reduce bud infestation with eggs and larvae. However, the mechanisms by which this occurs as well as the effects of other active substances are not known. In this study, the effects of the neonicotinoid insecticide Biscaya (a.i. thiacloprid) and the pyrethroids Mavrik (tau-fluvalinate) and Karate Zeon (lambda-cyhalothrin) applied at the bud stage of winter oilseed rape with recommended field rates on infestation of buds with eggs and larvae of pollen beetles were tested in field trials in Germany in 2013–2015. In additional greenhouse experiments, it was investigated whether effects on bud infestation were caused by lethal effects on pollen beetle or by insecticidal residues on plants causing sublethal effects. In the field trials, application of Biscaya and Mavrik significantly reduced the percentage of buds containing eggs and larvae in contrast to Karate Zeon. In 2014 and 2015, 14 days after application, bud infestation on the main raceme was reduced by 86 and 82%, respectively, in Biscaya-treated plots and by 51 and 71%, respectively, in Mavrik-treated plots compared to the untreated plots. In the greenhouse experiments, the lowest percentage of bud infestation with eggs and larvae was recorded on Biscaya-treated plants whereas on Mavrik-treated plants, there was no significant difference compared with the control. The results of the field trials show that Biscaya and Mavrik reduced oilseed rape bud infestation with eggs and larvae of pollen beetles primarily by lethal effects on overwintered pollen beetles or by repellency. However, Biscaya had additional effects on egg laying, which was supported by the greenhouse experiments.