Bovine herpesvirus 5 BICP0 complements the bovine herpesvirus 1 homolog

Bovine herpesvirus 1 (BoHV-1) and BoHV-5 are closely related (82% amino acid identity) but differ strongly in neuropathogenesis. The immediate-early gene for BICP0 is less conserved (70% amino acid identity) and may contribute to a dissimilar phenotype. A peculiar difference is a guanosine hexamer in the BICP0-1 gene which aligns with only five guanosines in the BICP0-5 gene and therefore results in a frameshift in the latter open reading frame. Thus, the C-terminal amino acid sequence (residues 643–676 of BICP0-1 vs. 655–720 of BICP0-5) is completely different. We introduced the BICP0-5 frameshift into the BoHV-1 genome cloned as a bacterial artificial chromosome (BoHV-1 BAC) using the Red recombination system with galK selection and counterselection. Transfection of MDBK cells with the resulting BAC produced recombinant virus that replicated like wild type BoHV-1 in vitro. Attempts to exchange the entire BICP0-1 gene by the BoHV-5 homolog using the same approach failed repeatedly. Therefore, we cotransfected purified BICP0⁻/galK⁺-BoHV-1 BAC DNA with a recombination plasmid coding for BICP0-5 with or without a HA tag into MDBK cells. BoHV-1 recombinants expressing the respective proteins were characterized. In vitro, all recombinants grew to similar titers as the parental viruses, which demonstrates that BICP0-5 compensates for the growth defect of BICP0⁻/galK⁺-BoHV-1 and functionally complements BICP0-1 of BoHV-1. We conclude that BICP0 may be suitable to positively select BoHV-1 recombinants with deletions or insertions of additional genes of interest.     

Remaining questions in the case for balanced harvesting

Balanced harvesting – harvesting all species and sizes in an ecosystem in proportion to their productivity – is a fisheries management strategy that has been suggested recently to increase yields, while reducing overall ecosystem impact. However, some aspects of balanced harvesting are controversial, including its call for extensive harvesting of juveniles and forage fish. Balanced harvesting also calls for targeting species and size‐classes that are not currently marketable, possibly at a significant economic cost. Some have argued that this cost is outweighed by the ecological benefits of maintaining the ecosystem size and trophic structures and by the benefits of extra yield for food security. There is broad consensus that balanced harvesting would require major changes to fishery management institutions and consumer behaviour, and it is unclear to what extent it is physically possible with current technologies. For this reason, we argue that steps to implement balanced harvesting are difficult to justify until the case for it is more clearly resolved. We outline some of the pivotal questions that must be answered to make a convincing case for or against balanced harvesting, many of which can be answered empirically. In identifying these questions, we hope to offer a constructive path forward in resolving some of the key issues in the balanced harvesting debate.     

Smart spatial incentives for market-based conservation

Market-based instruments such as payments, auctions or tradable permits have been proposed as flexible and cost-effective instruments for biodiversity conservation on private lands. Trading the service of conservation requires one to define a metric that determines the extent to which a conserved site adds to the regional conservation objective. Yet, while markets for conservation are widely discussed and increasingly applied, little research has been conducted on explicitly accounting for spatial ecological processes in the trading. In this paper, we use a coupled ecological-economic simulation model to examine how spatial connectivity may be considered in the financial incentives created by a market-based conservation scheme. Land use decisions, driven by changing conservation costs and the conservation market, are simulated by an agent-based model of land users. On top of that, a metapopulation model evaluates the conservational success of the market. We find that optimal spatial incentives for agents correlate with species characteristics such as the dispersal distance, but they also depend on the spatio-temporal distribution of conservation costs. We conclude that a combined analysis of ecological and socio-economic conditions should be applied when designing market instruments to protect biodiversity.     

Insect resistance in potatoes: sources,evolutionary relationships,morphological and chemical defenses,and ecogeographical associations

Summary The past 25 years, 1686 potato accessions, representing 100 species in the genus Solanum L., subgenus Potatoe, section Petota, were evaluated for field resistance to one or more of the following insect pests: green peach aphid, Myzus persicae (Sulzer); potato aphid, Macrosiphum euphorbiae (Thomas); Colorado potato beetle, Leptinotarsa decemlineata (Say); potato flea beetle, Epitrix cucumeris (Harris); and potato leafhopper, Empoasca fabae (Harris). Accessions highly resistant to green peach aphid were identified within 36 species, to potato aphid within 24 species, to Colorado potato beetle within 10 species, to potato flea beetle within 25 species, and to potato leafhopper within 39 species. Resistance levels were characteristic within Solanum species. Insect resistance appears to be a primitive trait in wild potatoes. Susceptibility was most common in the primitive and cultivated Tuberosa. Insect resistance was also characteristic of the most advanced species. The glycoalkaloid tomatine was associated with field resistance to Colorado potato beetle and potato leafhopper. Other glycoalkaloids were not associated with field resistance at the species level. Dense hairs were associated with resistance to green peach aphid, potato flea beetle, and potato leafhopper. Glandular trichomes were associated with field resistance to Colorado potato beetle, potato flea beetle, and potato leafhopper. Significant correlations between insect score and altitude of original collection were observed in six of thirteen species. Species from hot and arid areas were associated with resistance to Colorado potato beetle, potato flea beetle, and potato leafhopper. Species from cool or moist areas tended to be resistant to potato aphid.Abbreviations EBN Endosperm Balance Number     

Spatio-temporal dynamics of bacterial communities associated with two plant species differing in organic acid secretion: A one-year microcosm study on lupin and wheat

Plants are generally assumed to influence the surrounding soil microflora through rhizodeposition. However, the role of rhizodeposits, and especially organic acids, in structuring the bacterial communities is still poorly understood. In this study, we asked the question whether plants differing in organic acid secretion have a different impact on the soil bacterial communities, and if this is the case, to which extent this impact is due to different organic acid concentrations in the rhizosphere. To investigate this question, we compared white lupin and wheat. The former is a high organic acid-secreting species, while the latter secretes only low amounts of carboxylates. We grew the plants in large microcosms including root-free control compartments for one year (replanted every second month) and analyzed the spatio-temporal changes in soil ATP concentrations, as well as in diversity and structure of bacterial communities (using DNA- and RNA-based DGGE) along a root-soil gradient after two, six and twelve month's cultivation. Our results showed: i) that white lupin and wheat differed in their impact on soil ATP concentrations and on the structure of root bacterial communities; ii) that cultivation time was a key factor in explaining the observed differences in all the parameters studied; and iii) that the amounts of organic acids accounted for a significant proportion (15%) of the variability within root active communities. These results indicate that plants influence their associated bacterial communities in a species-specific way and that for communities living in the direct vicinity of roots (rhizoplane-endorhizosphere), a significant part of this influence can be attributed to root-secreted organic acids.     

Apigenin-induced suicidal erythrocyte death

Apigenin, a flavone in fruits and vegetables, stimulates apoptosis and thus counteracts cancerogenesis. Erythrocytes may similarly undergo suicidal cell death or eryptosis, characterized by cell shrinkage and phosphatidylserine exposure at the cell surface. Triggers of eryptosis include increase of cytosolic Ca(2+) activity ([Ca(2+)](i)), ceramide formation and ATP depletion. The present study explored the effect of apigenin on eryptosis. [Ca(2+)](i) was estimated from Fluo3-fluorescence, cell volume from forward scatter, phosphatidylserine exposure from annexin V binding, hemolysis from hemoglobin release, ceramide utilizing antibodies, and cytosolic ATP with luciferin-luciferase. A 48 h exposure to apigenin significantly increased [Ca(2+)](i) (≥ 1 μM), increased ceramide formation (15 μM), decreased ATP concentration (15 μM), decreased forward scatter (≥ 1 μM), and increased annexin V binding (≥ 5 μM) but did not significantly modify hemolysis. The effect of 15 μM apigenin on annexin V binding was blunted by Ca(2+) removal. The present observations reveal novel effects of apigenin, i.e. stimulation of Ca(2+) entry, ceramide formation and ATP depletion in erythrocytes with subsequent triggering of suicidal erythrocyte death, paralleled by cell shrinkage and phosphatidylserine exposure.     

Quantitative assessment of the fungal contribution to microbial tissue in soil

The fungi-to-bacteria ratio in soil ecological concepts and its application to explain the effects of land use changes have gained increasing attention over the past decade. Four different main approaches for quantifying the fungal and bacterial contribution to microbial tissue can be distinguished: (1) microscopic methods, (2) selective inhibition, (3) specific cell membrane components and (4) specific cell wall components. In this review, the different methods were compared and we hypothesized that all these approaches result in similar values for the fungal and bacterial contribution to total microbial biomass, activity, and residues (dead microbial tissue) if these methods are evenly reliable for the estimation of fungal biomass. The fungal contribution to the microbial biomass or respiration varied widely between 2 and 95% in different data sets published over the past three decades. However, the majority of the literature data indicated that fungi dominated microbial biomass, respiration or non-biomass microbial residues, with mean percentages obtained by the different methodological approaches varying between 35 and 76% in different soil groups, i.e. arable, grassland, and forest soils and litter layers. Microscopic methods generally gave the lowest average values, especially in arable and grasslands soils. Very low ratios in fungal biomass C-to-ergosterol obtained by microscopic methods suggest a severe underestimation of fungal biomass by certain stains. Relatively consistent ratios of ergosterol to linoleic acid (18:2ω6,9) indicate that both cell membrane components are useful indicators for saprotrophic and ectomycorrhizal fungi. More quantitative information on the PLFA content of soil bacteria and the 16:1ω5 content of arbuscular mycorrhizal fungi is urgently required to fully exploit the great potential of PLFA measurements. The most consistent results have been obtained from the analysis of fungal glucosamine and bacterial muramic acid in microbial residues. Component-specific δ¹³C analyses of PLFA and amino sugars are a promising prospect for the near future.     

Detection of exotic QTLs controlling nitrogen stress tolerance among wild barley introgression lines

Nitrogen (N) is one of the most important plant nutrients, controlling growth and, ultimately, yield of a cultivar. Hordeum vulgare ssp. spontaneum, the wild barley progenitor of cultivated barley, is known to possess genes that can improve tolerance against biotic and abiotic stresses. A quantitative trait locus (QTL) study with two levels of N fertilization was conducted under glasshouse in order to locate wild barley alleles that improve N stress tolerance in the genetic background of an elite barley cultivar. For this, a set of 28 barley introgression lines (S42ILs), which originate from the cross ??Scarlett???×???ISR42-8??, was studied. The S42ILs, containing single or multiple wild barley introgressions, and ??Scarlett?? were evaluated in regard to a total of 15 traits, related to morphological parameters, grain parameters as well as to carbon (C) and N content parameters. A mixed model analysis and a subsequent Dunnett test was conducted to identify S42ILs that significantly deviate from the recurrent parent ??Scarlett??, either tested separately for each N level, or simultaneously across both N levels. In total, 65 QTLs were detected for the S42IL set. Most QTLs were found for chlorophyll content during heading (10 QTLs) and the fewest for C/N ratio of straw (1 QTL). The individual S42ILs possessed different numbers of QTLs. For S42IL-108, a maximum of eight QTLs were found whereas S42IL-145 did not show any significant difference from ??Scarlett??. Wild barley alleles revealed decreasing effects at 32 QTLs and increasing effects at 33 QTLs. Although 25 QTLs exhibited similar effects across both N levels, 18 and 22 QTLs exhibited effects that were only detected under N0 or N1, respectively. We, thus, conclude that it may be worth to select improved barley cultivars for N stress tolerance separately under low N fertilization, rather than extrapolating trait performances from experiments carried out under standard N fertilization conditions. A number of wild barley QTL alleles improved N stress tolerance. For example, a wild barley QTL allele on chromosome 4H, present in the Hsp introgression of S42IL-119, was associated with a 13.0?% increase of thousand grain weight across both N levels and a 20?% increase under low N supply. QTLs detected in the present study were compared with those of previous field studies of the same cross and with other QTL studies in barley and other small grains. Accordance between QTL studies (QTLs showing similar effects at the same map location) is documented and discussed. Based on our study, promising wild barley QTL alleles are available in S42ILs, which can be readily utilized to select for improved N use efficiency in barley breeding.     

Studies on the aroma of five fresh tomato cultivars and the precursors of cis- and trans-4,5-epoxy-(E)-2-decenals and methional

Three tasty (BR-139, FA-624, and FA-612) and two less tasty (R-144 and R-175) fresh greenhouse tomato cultivars, which significantly differ in their flavor profiles, were screened for potent odorants using aroma extract dilution analysis (AEDA). On the basis of AEDA results, 19 volatiles were selected for quantification in those 5 cultivars using gas chromatography-mass spectrometry (GC-MS). Compounds such as 1-penten-3-one, ( E, E)- and ( E, Z)-2,4-decadienal, and 4-hydroxy-2,5-dimethyl-3(2 H)-furanone (Furaneol) had higher odor units in the more preferred cultivars, whereas methional, phenylacetaldehyde, 2-phenylethanol, or 2-isobutylthiazole had higher odor units in the less preferred cultivars. Simulation of the odor of the selected tomato cultivars by preparation of aroma models and comparison with the corresponding real samples confirmed that all important fresh tomato odorants were identified, that their concentrations were determined correctly in all five cultivars, and that differences in concentration, especially of the compounds mentioned above, make it possible to distinguish between them and are responsible for the differential preference. To help elucidate formation pathways of key odorants, labeled precursors were added to tomatoes. Biogenesis of cis- and trans-4,5-epoxy-( E)-2-decenals from linoleic acid and methional from methionine was confirmed.