The use of styrene-maleic acid copolymer (SMA) for studies on T cell membrane rafts

An emerging alternative to the use of detergents in biochemical studies on membrane proteins is apparently the use styrene-maleic acid (SMA) amphipathic copolymers. These cut the membrane into nanodiscs (SMA-lipid particles, SMALPs), which contain membrane proteins possibly surrounded by their native lipid environment. We examined this approach for studies on several types of T cell membrane proteins, previously defined as raft or non-raft associated, to see whether the properties of the raft derived SMALPs differ from non-raft SMALPs. Our results indicate that two types of raft proteins, GPI-anchored proteins and two Src family kinases, are markedly present in membrane fragments much larger (>250?nm) than those containing non-raft proteins (<20?nm). Lipid probes sensitive to membrane fluidity (membrane order) indicate that the lipid environment in the large SMALPs is less fluid (more ordered) than in the small ones which may indicate the presence of a more ordered lipid L_o phase which is characteristic of membrane rafts. Also the lipid composition of the small vs. large SMALPs is markedly different – the large ones are enriched in cholesterol and lipids containing saturated fatty acids. In addition, we confirm that T cell membrane proteins present in SMALPs can be readily immunoisolated. Our results support the use of SMA as a potentially better (less artifact prone) alternative to detergents for studies on membrane proteins and their complexes, including membrane rafts.     

Evolution of apomixis loci in Pilosella and Hieracium (Asteraceae) inferred from the conservation of apomixis-linked markers in natural and experimental populations

The Hieracium and Pilosella (Lactuceae, Asteraceae) genera of closely related hawkweeds contain species with two different modes of gametophytic apomixis (asexual seed formation). Both genera contain polyploid species, and in wild populations, sexual and apomictic species co-exist. Apomixis is known to co-exist with sexuality in apomictic Pilosella individuals, however, apomictic Hieracium have been regarded as obligate apomicts. Here, a developmental analysis of apomixis within 16 Hieracium species revealed meiosis and megaspore tetrad formation in 1 to 7% of ovules, for the first time indicating residual sexuality in this genus. Molecular markers linked to the two independent, dominant loci LOSS OF APOMEIOSIS (LOA) and LOSS OF PARTHENOGENESIS (LOP) controlling apomixis in Pilosella piloselloides subsp. praealta were screened across 20 phenotyped Hieracium individuals from natural populations, and 65 phenotyped Pilosella individuals from natural and experimental cross populations, to examine their conservation, inheritance and association with reproductive modes. All of the tested LOA and LOP-linked markers were absent in the 20 Hieracium samples irrespective of their reproductive mode. Within Pilosella, LOA and LOP-linked markers were essentially absent within the sexual plants, although they were not conserved in all apomictic individuals. Both loci appeared to be inherited independently, and evidence for additional genetic factors influencing quantitative expression of LOA and LOP was obtained. Collectively, these data suggest independent evolution of apomixis in Hieracium and Pilosella and are discussed with respect to current knowledge of the evolution of apomixis.     

Expression control of nitrile hydratase and amidase genes in Rhodococcus erythropolis and substrate specificities of the enzymes

Bacterial amidases and nitrile hydratases can be used for the synthesis of various intermediates and products in the chemical and pharmaceutical industries and for the bioremediation of toxic pollutants. The aim of this study was to analyze the expression of the amidase and nitrile hydratase genes of Rhodococcus erythropolis and test the stereospecific nitrile hydratase and amidase activities on chiral cyanohydrins. The nucleotide sequences of the gene clusters containing the oxd (aldoxime dehydratase), ami (amidase), nha1, nha2 (subunits of the nitrile hydratase), nhr1, nhr2, nhr3 and nhr4 (putative regulatory proteins) genes of two R. erythropolis strains, A4 and CCM2595, were determined. All genes of both of the clusters are transcribed in the same direction. RT-PCR analysis, primer extension and promoter fusions with the gfp reporter gene showed that the ami, nha1 and nha2 genes of R. erythropolis A4 form an operon transcribed from the Pami promoter and an internal Pnha promoter. The activity of Pami was found to be weakly induced when the cells grew in the presence of acetonitrile, whereas the Pnha promoter was moderately induced by both the acetonitrile or acetamide used instead of the inorganic nitrogen source. However, R. erythropolis A4 cells showed no increase in amidase and nitrile hydratase activities in the presence of acetamide or acetonitrile in the medium. R. erythropolis A4 nitrile hydratase and amidase were found to be effective at hydrolysing cyanohydrins and 2-hydroxyamides, respectively.     

Benthic macroinvertebrates along the Czech part of the Labe and lower section of the Vltava rivers from 1996–2005, with a particular focus on rare and alien species

In the Czech part of the Labe River and the lower part of the Vltava River, we examined if the benthic macroinvertebrate composition changed from 1996 to 2005 due to expected improvements in water quality resulting from socioeconomic changes in the Czech Republic since the 1990s. Special attention was given to rare and alien species. The four biological metrics used (Number of taxa, BMWP, Number of sensitive taxa, and Number of EPT taxa) demonstrated that there was indeed an improvement in water quality as well as a slight improvement of the Labe microhabitats during the investigated period. An increasing Number of taxa over time was observed at most sites. Two main concurrent ecological processes are recently in progress in the Labe: a recovery of native species and an expansion of alien species, some of which are considered invasive. The caddisfly Setodes punctatus and the beetle Pomatinus substriatus, considered as regionally extinct in the Czech Republic until 2005, were rediscovered during our investigations. Findings of the crustacean Hemimysis anomala (invasive) and the chironomids Stenochironomus sp. and Lipiniella sp. were the first records of these taxa in the Czech Republic.     

The meek inherit the earth: less aggressive wild mice are more successful in challenging situations

Numerous studies have shown an association between aggressiveness and several other behavioural traits. For example, more aggressive animals were bold and active explorers tending to form persistent routines whereas less aggressive animals were shy, careful but more flexible. While the former are thought to be more successful under stable conditions the latter should have advantages in more dynamic situations. These differences can apply not only to individuals but also to populations, species or groups of species with important implications to species distributions and speciation rates. Here we utilized the Morris water task (MWT) to investigate how two subspecies, Mus musculus musculus and M. m. domesticus, known to differ in aggressiveness, cope with stressful situations. We found that less aggressive musculus males performed significantly better in solving the MWT than more aggressive domesticus males. This suggests that M. m. musculus is more flexible and could be more successful under stressful and/or dynamic situations typical of dispersal bouts. It seems plausible that this difference may have had an influence on the secondary contact between musculus and domesticus populations in the past and perhaps still can affect the dynamics of the European hybrid zone between the subspecies. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113 , 310–319.     

Resurrecting Helix straminea,a forgotten escargot with trans‐Adriatic distribution: first insights into the genetic variation within the genus Helix (Gastropoda: Pulmonata)

Helix is a genus of large Western Palaearctic land snails, particularly diverse in the Mediterranean region. Despite the large size and attractiveness of its members, it has an unsettled taxonomy, and no data are available on its intrageneric phylogenetic relationships. One of the problematic Helix taxa is the widely distributed, economically important, and conchologically very variable H. lucorum. Two distinct forms may be encountered under this name in the Apennine Peninsula: a typical one in the north, and a form originally described as Helix straminea in central and southern Italy. To evaluate the status of H. straminea and its relationships to Italian and Balkan Helix fauna, we combined shell morphology, geometric morphometrics, and phylogenetic analysis based on two mitochondrial genes. Distribution data were improved by drawing information from unambiguously identifiable photographs posted online. Based on our results, Helix straminea is redescribed as a separate species, and we find it to have a disjunctive trans‐Adriatic range with closest relatives in the western Balkans. We provide insight into relationships and intraspecific variability within Helix, a first step towards a comprehensive revision of the genus. On the example of Italian Helix fauna, we demonstrate how understanding of snail zoogeography may change with improving taxonomy. © 2014 The Linnean Society of London     

Signalling mechanisms involved in the response of two varieties of Humulus lupulus L. to soil drying: I. changes in xylem sap pH and the concentrations of abscisic acid and anions

Background and aims

Soil drying leads to the generation of chemical signals in plants that regulate water use via control of the stomatal aperture. The aim of our work was to identify the presence and identity of potential chemical signals, their dynamics, and their relationship with transpiration rate during soil drying in hop (Humulus lupulus (L.)) plants.

Methods

We used pressure chamber technique for measurement of shoot water potential and collection of shoot xylem sap. We analyzed concentrations of abscisic acid (ABA), nitrate, phosphate, sulphate and malate in sap and also the rate of whole plant transpiration.

Results

Transpiration rate decreased prior to changes in shoot water potential. The concentration of ABA in xylem sap continuously increased from early to later stages of water stress, whereas in leaves it increased only at later stages. Shoot sap pH increased simultaneously with the decrease of transpiration rate. Xylem sap alkalization was in some cases accompanied by a decrease in nitrate concentration and an increase in malate concentration. Concentration of sulphate increased in xylem sap during drying and sulphate in combination with a higher ABA concentration enhanced stomatal closure.

Conclusions

Several early chemical signals appear in sap of hop plants during soil drying and their impact on transpiration may vary according to the stage of soil drying.