Comparative proteomics of Dehalococcoides spp. reveals strain-specific peptides associated with activity

Anaerobic reductive dehalogenation by Dehalococcoides spp. is an ideal system for studying functional diversity of closely related strains of bacteria. In Dehalococcoides spp., reductive dehalogenases (RDases) are key respiratory enzymes involved in the anaerobic detoxification of halogenated compounds at contaminated sites globally. Although housekeeping genes sequenced from Dehalococcoides spp. are >85% identical at the amino acid level, different strains are capable of dehalogenating diverse ranges of compounds, depending largely on the suite of RDase genes that each strain harbors and expresses. We identified RDase proteins that corresponded to known functions in four characterized cultures and predicted functions in an uncharacterized Dehalococcoides-containing mixed culture. Homologues within RDase subclusters containing PceA, TceA, and VcrA were among the most frequently identified proteins. Several additional proteins, including a formate dehydrogenase-like protein (Fdh), had high coverage in all strains and under all growth conditions.     

Effect of weather conditions on the flight activity of Nigerian blackflies (Diptera: Simuliidae)

Abstract. A vehicle-mounted net was used to make hourly catches of blackflies at 700–1000 m altitude in the River Assob valley, central Nigeria, on 20 days during the dry season. Pearson Correlation Matrix analysis of the collection data and meteorology showed that the main factors affecting flight activity of each of the four most abundant blackfly species were primarily light intensity and secondly wind velocity, whereas relative humidity was the least important factor. Right activity showed a negative linear regression against wind, with some activity occurring in wind speeds up to 15 km/h. Distance-weighted least-squares (DWLS) regressions showed little correlation of temperature with activity for Simulium hargreavesi and S.adersi , but S.squamosum and S.vorax had small peaks at 28C and 31C, respectively. DWLS regression against light intensity showed an activity peak at 6000 lux, except in Simulium adersi.     

An ATPase activity associated with brain microtubules.

A persistent ATPase/GTPase activity has been found to be associated with highly recycled bovine brain microtubules. A GTP regeneration system was introduced to minimize the inhibitory effects of this hydrolase on microtubule polymerization. The characteristics of the ATPase indicate that it is not involved in GTP-induced mictrotubule polymerization, but is directly involved in ATP-induced polymerization. ATP-induced polymerization was also shown to require stoichiometric amounts of GDP, but higher levels of GDP inhibited both microtubule formation and the ATPase activity. An ammonium sulfate fractionation procedure was devised to separate microtubule protein into an ATPase-rich fraction and a pure tubulin fraction. The pure tubulin fraction polymerized in the presence of GTP, but not in the presence of ATP and GDP. In contrast, the ATPase-rich fraction polymerized with either ATP or GTP. It is still not known whether the microtubule associated ATPase plays a significant role in cellular microtubule function.     

Characterization of the anti-fungal activity of a Bacillus spp. associated with sclerotia from Sclerotinia sclerotiorum

Sclerotinia sclerotiorum fruiting bodies (sclerotia) were found to harbour bacteria that possess anti-fungal activity. Among 1,140 bacterial isolates collected, 32 were found to inhibit the growth of four common fungal pathogens of canola, S. sclerotiorum, Rhizoctonia solani, Alternaria brassicae and Leptosphaeria maculans. One of these broad-spectrum isolates, LEV-006, was found to be closely related to Bacillus subtilis based on 16S rRNA analysis. The anti-fungal activities were purified and found to be associated with a low molecular weight peptide complex consisting mostly of the cyclic lipopeptide fengycin A and B, as revealed by matrix-assisted laser desorption/ionization time-of-flight and post-source decay analysis, as well as two proteins of 20 and 55 kDa. Peptide mass fingerprinting revealed that the 55-kDa protein was similar to vegetative catalase 1; however, when the enzyme was expressed in Escherichia coli, it exhibited catalase but not anti-fungal activity. The sequences of several peptides from the 20-kDa protein were obtained and indicated that it was a unique anti-fungal protein.     

Influence of weather conditions on the flight of migrating black storks

This study tested the potential influence of meteorological parameters (temperature, humidity, wind direction, thermal convection) on different migration characteristics (namely flight speed, altitude and direction and daily distance) in 16 black storks (Ciconia nigra). The birds were tracked by satellite during their entire autumnal and spring migration, from 1998 to 2006. Our data reveal that during their 27-day-long migration between Europe and Africa (mean distance of 4100 km), the periods of maximum flight activity corresponded to periods of maximum thermal energy, underlining the importance of atmospheric thermal convection in the migratory flight of the black stork. In some cases, tailwind was recorded at the same altitude and position as the birds, and was associated with a significant rise in flight speed, but wind often produced a side azimuth along the birds'' migratory route. Whatever the season, the distance travelled daily was on average shorter in Europe than in Africa, with values of 200 and 270 km d⁻¹, respectively. The fastest instantaneous flight speeds of up to 112 km h⁻¹ were also observed above Africa. This observation confirms the hypothesis of thermal-dependant flight behaviour, and also reveals differences in flight costs between Europe and Africa. Furthermore, differences in food availability, a crucial factor for black storks during their flight between Europe and Africa, may also contribute to the above-mentioned shift in daily flight speeds.     

Productive infection of HUVEC by HHV-8 is associated with changes compatible with angiogenic transformations

Kaposi's Sarcoma (KS) is an angioproliferative disease associated with human herpesvirus 8 (HHV-8) infection. We have characterized the morphologic and phenotypic modifications of HUVEC in a model of productive HHV-8 infection. HHV-8 replication was associated with ultra-structural changes, flattened soma and a loss of marginal folds and intercellular contacts, and morphologic features, spindle cell conversion and cordon-like structures formation. Phenotypic changes observed on cordon-like structures included partial loss and redistribution of CD31/PECAM-1 and VE-cadherin, uPAR up-regulation and de novo expression of CD13/APN. Such changes demonstrate the induction, in HUVEC, of an angiogenic profile. Most of these findings are directly linked to HHV-8-encoded proteins expression, suggesting that HHV-8 itself may participate to the initial steps of the angiogenic transformation in KS.     

Genomic changes associated with somaclonal variation in banana (Musa spp.)

The molecular basis of somaclonal variation is not precisely known, but both genetic and epigenetic mechanisms have been proposed. The available evidence points toward the existence of labile portions of the genome that can be modulated when the cells undergo the stress of tissue culture. Therefore, the hypothesis that there are identifiable and predictable DNA markers for the early diagnosis of somaclonal variation has been tested. Representational difference analysis was used to isolate unique fragments of DNA (difference products) between visible culture-induced off-type and normal banana plants. Markers generated from six difference products differentiated between some of the off-type and normal pairs. The genomic region around one of these difference products has been extensively characterized and has a high degree of polymorphism, with variation in up to 10% of the nucleotides sequenced in the region. This same region has been shown to vary in other pairs of off-type and normal banana plants derived from tissue culture as well as in plants propagated commercially in vitro. The data are consistent with the hypothesis that there is at least one particularly labile portion of the genome that is especially susceptible to the stress imposed during tissue culture and that is associated with higher rearrangement and mutation rates than other portions of the genome. Consequently, the regions that are reported here have the potential to be used as early detection tools for identifying somaclonal variants.     

Light-induced enzyme activity changes associated with the photoisomerization of bound spiropyran

A spiropyran compound which undergoes photoisomerization was covalently attached to several enzymes in order to alter their activity by imposing light on them. The enzyme-bound spiropyran exhibited normal photochromism when the matrix enzyme was hydrophobic in nature, whereas reverse photochromism was evident when the spiropyran was attached to hydrophilic enzymes. The activity of the modified enzymes was found to change because of photoisomerization of the bound spiropyran. A fluorescent probe for hydrophobic sites revealed that the modified enzymes remained hydrophobic when the bound spiropyran closed and turned hydrophilic when the spiropyran opened. A mechanism for the photoinduced activity changes is postulated in relation to the enzyme-substrate affinity in the open- and closed-ring states of spiropyran.     

An electron microscopic investigation of age-dependent changes in the flight muscle of Musca domestica L.

The thoracic flight muscle from adult male NAIDM house flies was examined, from emergence to very old age. Thin sections stained with uranyl acetate and bismuth showed no myofibrillar or mitochondrial degeneration from 1 day to 19 days post-emergence, contrary to earlier reports. Some progressive loss in glycogen content and increase in mitochondrial size were observed for muscle from young to very old flies. However, there was no conclusive evidence of fusion of smaller mitochondria into larger ones with advancing age, despite exhaustive examination of representative sections of muscle samples of adult males of different ages.     

Bacterial communities associated with tuberculate ectomycorrhizae of Rhizopogon spp.

We have previously reported the design of a new PCR primer pair that allows amplification of a broad range of eubacterial 16S rDNA sequences from ectomycorrhizae (ECM) without co-amplification of plastid or mitochondrial sequences. Here, we report using a similar primer combination to generate three small 16S rDNA libraries from tuberculate ECM of Rhizopogon spp., two from R. vinicolor ECM (libraries Rvi18 and Rvi24) and one from R. vesiculosus ECM (library Rve13). At the class level, libraries were dominated by sequences from the Alphaproteobacteria, Gammaproteobacteria, and Acidobacteria, with some Sphingobacteria, Actinobacteria, Planctomycetacia, and Verrucomicrobiae present as well. Based on the parsimony test implemented in TreeClimber, libraries Rvi18 and Rvi24 were significantly different from Rve13 at the α = 0.05 level, while they were only borderline significantly different from each other (p = 0.07). Differences between Rvi and Rve libraries were primarily due to differences in the number of Alphaproteobacteria sequences and specifically sequences from the Rhizobiales, which were more common in the Rve13 library. It is currently unknown what drives these differences between eubacterial communities. Amplification success for eubacterial 16S rDNA sequences was generally low in this study indicating low abundance of bacteria on tuberculate ECM. Attempts to amplify nitrogenase reductase (nifH) sequences were unsuccessful. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Biological control of Orobanche spp. with Phytomyza orobanchia, a review

This review summarises theavailable information on the biocontrol agentPhytomyza orobanchia Kalt. (Diptera, Agromyzidae). It gives an overview of a rareexample of weed biocontrol using insects in aninundative approach. A high diversity ofphytophagous insects has been collected onparasitic weeds of the genus Orobanche(Orobanchaceae). For biological control of Orobanche spp., only insects like P. orobanchia whose host range is restricted toOrobanche spp. are of interest. Of the140 Orobanche spp. described in total,P. orobanchia is reported from 21 species. The larvae of P. orobanchia minein Orobanche shoots and capsules. As aconsequence, a natural reduction of Orobancheseed production by 30 to almost 80%has been reported from different countries. Theefficacy of P. orobanchia under naturalconditions is limited by low temperatures, cultural practices and natural enemies. Tostrengthen the natural population and itsimpact, inundative releases of P. orobanchia adults at the beginning of Orobanche emergence have to be undertaken. Different methods for the application of P. orobanchia in biocontrol of Orobanche spp. have been developed in theformer Soviet Union. Releases of 500 to 1000adults/ha resulted in a reduction of up to 96%of the Orobanche seed production.However, due to the 10 to 15 year longevity ofOrobanche seeds, further infestationswill occur in the following cropping seasons.Consequently, releases of P. orobanchiahave to be repeated continuously over severalyears to reduce the infestation to a tolerablelevel.     

The influence of weather on western corn rootworm flight activity at the borders of a soybean field in east central Illinois

The evolution of a new strain of western corn rootworm (WCR) beetle (Diabrotica virgifera virgifera) that has adapted to crop rotation by flying from host cornfields to nearby soybean fields to lay eggs is presenting new challenges to farmers in the U.S.A. Corn Belt. Development of effective management tools for the WCR that display this new behavior require knowledge of atmospheric factors that influence their interfield movement. In this study, WCR movement into a soybean field is related to solar radiation, wind speed and direction, air temperature, and precipitation with consideration of biological factors that also influence flight. WCR flight activity and meteorological variables were measured above the canopy of a 1.64 ha soybean field in east-central Illinois between late July and early September, 1997. On 14 days, insect traps were sampled at 30-min intervals. Interfield movement of WCR occurred over a wide range of solar irradiances, air temperatures, and wind speeds. Darkness, air temperatures below 15 °C or above 31 °C, and wind speeds in excess of 2.0 m s^{- 1} prohibited aerial movement of WCR. Within these limits, atmospheric factors had only little influence on the biologically-driven temporal patterns of seasonal and daily WCR flight activity. Atmospheric conditions were conducive to WCR flight 62% of the time during the growing season when WCR were active. Weather conditions substantially reduced interfield WCR movement throughout about one-third of the days when female WCR beetles were abundant at the study site in 1997.     

沙棘属植物弗兰克氏菌研究进展

弗兰克氏菌(Frankia spp.)能够与沙棘等非豆科植物形成根瘤进行共生固氮,其固氮效率远远高于豆科植物根瘤菌,与沙棘共生的弗兰克氏菌还能够促进沙棘对旱寒等各种不同生境的适应性,是自然界一类具有开发潜力的放线菌资源。为了更好地开发利用弗兰克氏菌资源,推进弗兰克氏菌分类鉴定工作,加强弗兰克氏菌与寄主植物共生结瘤固氮的机制研究,促使弗兰克氏菌在农业生产中得到尽快应用,本文简要介绍沙棘属(Hippophae L.)物种多样性、结瘤状况与分布特点、沙棘根瘤形态结构与功能、弗兰克氏菌物种多样性与分布特征,讨论弗兰克氏菌的结瘤机制、生理生态效应与作用机制以及影响沙棘属植物与弗兰克氏菌共生的主要因子,以期为进一步开展沙棘属植物弗兰克氏菌的系统研究提供有价值的参考。     

The effect of wind-borne odours on the direction of flight in tsetse flies, Glossina spp.

ABSTRACT. The direction of flight in tsetse flies ( Glossina pallidipes Aust. and G. m. morsitans Westw.) taking off in the presence of certain wind-borne odours showed a significant upwind shift both in the field and in the laboratory. The average angular deviation between the resting orientation and flight direction was not materially affected by odour, but turns were steered in relation to wind direction if odour was present. Upwind flight in an odour plume was regularly preceded by a standing turn, the fly turning partly or completely into the wind before taking off in upwind flight. This suggests that wind direction was assessed, and flight direction determined, before the fly took off.