Nocturnal windborne migration of ground beetles,particularly Pseudoophonus griseus (Coleoptera: Carabidae), in China

1 A network of light‐traps, an aerial net carried by kytoon (balloon) and two entomological radars were used to investigate whether ground beetles migrate nocturnally through China. The network‐wide, simultaneous sudden increase in light trap catches, and after subsequent decrease, indicated a seasonal long‐distance night migration of ground beetles, with Pseudoophonus griseus (Panzer) predominant, in August. 2 Aerial net trapping indicated that carabids were able to ascend to altitudes of at least 200 m and become windborne. Radar observations indicated that the migratory beetles formed high‐density layer concentrations at approximately 200–300 m. 3 These concentrations were coincident with the top of the temperature inversion and a wind speed maximum, which suggested that the carabids tended to select warm, fast moving air for their long‐distance migration. 4 The ground beetles orientated and displaced towards the downwind direction in southerly winds. Their air speed decreased as the tailwind increased and, thus, migrating beetles appeared to be conserving energy. 5 The mean ± SD displacement speed (ground speed) and air speed were 6.85 ± 1.73 m/s (n= 172) and 4.45 ± 1.54 m/s (n= 172), respectively. The duration of flight, estimated from the variation in area density derived from radar data, was approximately 9–10 h, indicating that the beetles might migrate hundreds of kilometres in a single flight.     

Intracellular phosphorus metabolism of Microcystis aeruginosa under various redox potential in darkness

Phosphorus metabolism of Microcystis aeruginosa was studied under gradient redox potential from 252 mV to –70 mV in darkness. The release of phosphorus occurred in all the treatments, and this process was accelerated in darkness when the redox potential was lowered. Low redox potential in darkness stimulated the accumulation of polyphosphate (PolyP) and the degradation of polyglucose. The synthesis of PolyP delayed the decrease of intracellular orthophosphate. The death of M. aeruginosa was slowered when the redox potential was low in darkness. The accumulation of PolyP under low redox potential in the dark was very important to M. aeruginosa for endurance through the unfavorable growth conditions for maintaining phosphorus concentration, energy storage, and other physiological functions. The ability to accumulate PolyP in the dark and negative redox potential may be of considerable advantage in the low-light, organically rich, and low-redox habitats.     

Structure and functional analysis of the microbial community in an aerobic: anaerobic sequencing batch reactor (SBR) with no phosphorus removal

The bacterial community of an aerobic:anaerobic non-P removing SBR biomass fed a mixture of acetate and glucose was analysed using several 16S rRNA based methods. Populations responsible for anaerobic glucose and acetate assimilation were determined with fluorescent in situ hybridization (FISH) in combination with microautoradiography (FISH/MAR). At 'steady state' this community consisted of alpha-Proteobacteria (26%) and gamma-Proteobacteria (14%), mainly appearing as large cocci in tetrads (i.e. typical 'G-Bacteria'). Large numbers of low G+C bacteria (22%), and high G+C Gram-positive bacteria (29%) seen as small cocci in clusters or in sheets were also detected after FISH. DGGE fingerprinting of PCR amplified 16S rDNA fragments and subsequent cloning and sequencing of several of the major bands led to the identification of some of these populations. They included an organism 98% similar in its 16S rRNA sequence to Micropruina glycogenica, and ca. 76% of the high G+C bacteria responded to a probe MIC 184, designed against it. The rest responded to the KSB 531 probe designed against a high G+C clone sequence, sbr-gs28 reported in other similar systems. FISH analyses showed that both these high G+C populations were almost totally dominated by small clustered cocci. Only ca. 2% of cells were beta-Proteobacteria. None of the alpha- and gamma-Proteobacterial 'G-bacteria' responded to FISH probes designed for the 'G-Bacteria' Amaricoccus spp. or Defluvicoccus vanus. FISH/MAR revealed that not all the alpha-Proteobacterial 'G-Bacteria' could take up acetate or glucose anaerobically. Almost all of the gamma-Proteobacterial 'G-Bacteria' assimilated acetate anaerobically but not glucose, the low G+C clustered cocci only took up glucose, whereas the high G+C bacteria including M. glycogenica and the sbr-gs28 clone assimilated both acetate and glucose. All bacteria other than the low G+C small cocci and a few of the alpha-Proteobacteria accumulated PHB. The low G+C bacteria showing anaerobic glucose assimilation ability were considered responsible for the lactic acid produced anaerobically by this SBR biomass, and M. glycogenica for its high glycogen content.     

Monitoring Rates and Heterogeneity of High-Pressure Germination of Bacillus Spores by Phase-Contrast Microscopy of Individual Spores

Germination of Bacillus spores with a high pressure (HP) of ∼150 MPa is via activation of spores'' germinant receptors (GRs). The HP germination of multiple individual Bacillus subtilis spores in a diamond anvil cell (DAC) was monitored with phase-contrast microscopy. Major conclusions were that (i) >95% of wild-type spores germinated in 40 min in a DAC at ∼150 MPa and 37°C but individual spores'' germination kinetics were heterogeneous; (ii) individual spores'' HP germination kinetic parameters were similar to those of nutrient-triggered germination with a variable lag time (T_lag) prior to a period of the rapid release (ΔT_release) of the spores'' dipicolinic acid in a 1:1 chelate with Ca²⁺ (CaDPA); (iii) spore germination at 50 MPa had longer average T_lag values than that at ∼150 MPa, but the ΔT_release values at the two pressures were identical and HPs of <10 MPa did not induce germination; (iv) B. subtilis spores that lacked the cortex-lytic enzyme CwlJ and that were germinated with an HP of 150 MPa exhibited average ΔT_release values ∼15-fold longer than those for wild-type spores, but the two types of spores exhibited similar average T_lag values; and (v) the germination of wild-type spores given a ≥30-s 140-MPa HP pulse followed by a constant pressure of 1 MPa was the same as that of spores exposed to a constant pressure of 140 MPa that was continued for ≥35 min; (vi) however, after short 150-MPa HP pulses and incubation at 0.1 MPa (ambient pressure), spore germination stopped 5 to 10 min after the HP was released. These results suggest that an HP of ∼150 MPa for ≤30 s is sufficient to fully activate spores'' GRs, which remain activated at 1 MPa but can deactivate at ambient pressure.     

Monitoring bacterial growth using tunable resistive pulse sensing with a pore-based technique

A novel bacterial growth monitoring method using a tunable resistive pulse sensor (TRPS) system is introduced in this study for accurate and sensitive measurement of cell size and cell concentration simultaneously. Two model bacterial strains, Bacillus subtilis str.168 (BSU168) and Escherichia coli str.DH5α (DH5α), were chosen for benchmarking the growth-monitoring performance of the system. Results showed that the technique of TRPS is sensitive and accurate relative to widely used methods, with a lower detection limit of cell concentration measurement of 5?×?10⁵ cells/ml; at the same time, the mean coefficient of variation from TRPS was within 2 %. The growth of BSU168 and DH5α in liquid cultures was studied by TRPS, optical density (OD), and colony plating. Compared to OD measurement, TRPS-measured concentration correlates better with colony plating (R?=?0.85 vs. R?=?0.72), which is often regarded as the gold standard of cell concentration determination. General agreement was also observed by comparing TRPS-derived cell volume measurements and those determined from microscopy. We have demonstrated that TRPS is a reliable method for bacterial growth monitoring, where the study of both cell volume and cell concentration are needed to provide further details about the physical aspects of cell dynamics in real time.     

Heterologous overexpression of Vigna radiata epoxide hydrolase in Escherichia coli and its catalytic performance in enantioconvergent hydrolysis of p-nitrostyrene oxide into (R)-p-nitrophenyl glycol

Two native epoxide hydrolases (EHs) were previously discovered from mung bean powder (Vigna radiata), both of which can catalyze the enantioconvergent hydrolysis of p-nitrostyrene oxide (pNSO). In this study, the encoding gene of VrEH1 was successfully cloned from the cDNA of V. radiata by RT-PCR and rapid amplification of cDNA ends (RACE) technologies. High homologies were found to two putative EHs originated from Glycine max (80 %) and Medicago truncatula (79 %). The vreh1 gene constructed in pET28a(+) vector was then heterologously overexpressed in Escherichia coli BL21(DE3), and the encoded protein was purified to homogeneity by nickel affinity chromatography. It was shown that VrEH1 has an optimum activity at 45 °C and is very thermostable with an inactivation energy of 468 kJ mol^-1. The enzyme has no apparent requirement of metal ions for activity, and its activity was strongly inhibited by 1 mM of Ni²⁺, Cu²⁺, Fe²⁺, or Co²⁺. By adding 0.1 % Triton X-100, the enzyme activity could be significantly increased up to 340 %. VrEH1 shows an unusual ability of enantioconvergent catalysis for the hydrolysis of racemic pNSO, affording (R)-p-nitrophenyl glycol (pNPG). It displays opposite regioselectivity toward (S)-pNSO (83 % to C_α) in contrast to (R)-pNSO (87 % to C_β). The K _M and k _cat of VrEH1 were determined to be 1.4 mM and 0.42 s^-1 for (R)-pNSO and 5.5 mM and 6.2 s^-1 for (S)-pNSO. This thermostable recombinant VrEH1 with enantioconvergency is considered to be a promising biocatalyst for the highly productive preparation of enantiopure vicinal diols and also a good model for understanding the mechanism of EH stereoselectivity.     

Microtubule dynamics is required for root elongation growth under osmotic stress in Arabidopsis

Osmotic stress caused by drought and soil salinity is one of the factors that affect plant root system growth and development. Previous studies have shown that microtubule plays a critical role in plant roots response to osmotic stress, however, the underlying mechanism remains unclear. In the present study, the microtubule orientations in Arabidopsis roots growing under osmotic stress were determined using confocal fluorescence microscopy. The results showed that osmotic stress could significantly inhibit primary root elongation in Arabidopsis, and pharmacological tests confirmed that microtubules were involved in Arabidopsis roots response to osmotic stress. In vivo visualization of microtubule structures with the microtubule-binding domain–green fluorescent protein (GFP) reporter revealed altered microtubule orientation in rhizodermal cells under osmotic stress. These results above indicated that osmotic stress could inhibit the elongation growth of Arabidopsis primary root, and the inhibition effects might result from the changes in microtubule orientation.     

Toxic effects of nanoparticles on bioluminescence activity,seed germination,and gene mutation

The potential environmental toxicities of several metal oxide nanoparticles (NPs; CuO, TiO₂, NiO, Fe₂O₃, ZnO, and Co₃O₄) were evaluated in the context of bioluminescence activity, seed germination, and bacterial gene mutation. The bioassays exhibited different sensitivities, i.e., each kind of NP exhibited a different level of toxicity in each of the bioassays. However, with a few exceptions, CuO and ZnO NPs had most toxic for germination of Lactuca seed (EC₅₀ 0.46 mg CuO/l) and bioluminescence (EC₅₀ 1.05 mg ZnO/l). Three NPs (Co₃O₄, TiO₂, and Fe₂O₃) among all tested concentrations (max. 1,000 mg/l) showed no inhibitory effects on the tested organisms, except for Co₃O₄ NPs on bioluminescence activity (EC₅₀ 62.04 mg/l). The sensitivity of Lactuca seeds was greater than that of Raphanus seeds (EC₅₀ 0.46 mg CuO/l versus 26.84 mg CuO /l ). The ranking of metal toxicity levels on bioluminescence was in the order of ZnO?>?CuO?>?Co₃O₄?>?NiO?>?Fe₂O₃, TiO₂, while CuO?>?ZnO?>?NiO?>?Co₃O₄, Fe₂O₃, TiO₂ on germination. No revertant mutagenic ratio (greater than 2.0) of Salmonella typhimurium TA 98 was observed under any tested condition. These findings demonstrate that several bioassays, as opposed to any single one, are needed for the accurate assessment of NP toxicity on ecosystems.