Isolation, characterization and electron microscopic single particle analysis of the NADH : ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus

The proton-translocating NADH:ubiquinone oxidoreductase (complex I) has been purified from Aquifex aeolicus, a hyperthermophilic eubacterium of known genome sequence. The purified detergent solubilized enzyme is highly active above 50 degreesC. The specific activity for electron transfer from NADH to decylubiquinone is 29 U/mg at 80 degreesC. The A. aeolicus complex I is completely sensitive to rotenone and 2-n-decyl-quinazoline-4-yl-amine. SDS polyacrylamide gel electrophoresis shows that it may contain up to 14 subunits. N-terminal amino acid sequencing of the bands indicates the presence of a stable subcomplex, which is composed of subunits E, F, and G. The isolated complex is highly stable and active in a temperature range from 50 to 90 degreesC, with a half-life of about 10 h at 80 degreesC. The activity shows a linear Arrhenius plot at 50-85 degreesC with an activation energy at 31.92 J/mol K. Single particle electron microscopy shows that the A. aeolicus complex I has the typical L-shape. However, visual inspection of averaged images reveals many more details in the external arm of the complex than has been observed for complex I from other sources. In addition, the angle (90degrees) between the cytoplasmic peripheral arm and the membrane intrinsic arm of the complex appears to be invariant.

Particle-attached and free-living bacterial communities in a contrasting marine environment: Victoria Harbor, Hong Kong

Diversity of particle-attached and free-living marine bacteria in Victoria Harbor, Hong Kong, and its adjacent coastal and estuarial environments was investigated using DNA fingerprinting and clone library analysis. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes showed that bacterial communities in three stations of Victoria Harbor were similar, but differed from those in adjacent coastal and estuarine stations. Particle-attached and free-living bacterial community composition differed in the Victoria Harbor area. DNA sequencing of 28 bands from DGGE gel showed Alphaproteobacteria was the most abundant group, followed by the Bacteroidetes, and other Proteobacteria. Bacterial species richness (number of DGGE bands) differed among stations and populations (particle-attached and free-living; bottom and surface). BIOENV analysis indicated that the concentrations of suspended solids were the major contributing parameter for the spatial variation of total bacterial community structure. Samples from representative stations were selected for clone library (548 clones) construction and their phylogenetic distributions were similar to those of sequences from DGGE. Approximately 80% of clones were affiliated to Proteobacteria, Bacteroidetes and Cyanobacteria. The possible influences of dynamic pollution and hydrological conditions in the Victoria Harbor area on the particle-attached and free-living bacterial community structures were discussed.

Particle-attached and free-living bacterial communities in a contrasting marine environment: Victoria Harbor, Hong Kong

Diversity of particle-attached and free-living marine bacteria in Victoria Harbor, Hong Kong, and its adjacent coastal and estuarial environments was investigated using DNA fingerprinting and clone library analysis. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes showed that bacterial communities in three stations of Victoria Harbor were similar, but differed from those in adjacent coastal and estuarine stations. Particle-attached and free-living bacterial community composition differed in the Victoria Harbor area. DNA sequencing of 28 bands from DGGE gel showed Alphaproteobacteria was the most abundant group, followed by the Bacteroidetes, and other Proteobacteria. Bacterial species richness (number of DGGE bands) differed among stations and populations (particle-attached and free-living; bottom and surface). BIOENV analysis indicated that the concentrations of suspended solids were the major contributing parameter for the spatial variation of total bacterial community structure. Samples from representative stations were selected for clone library (548 clones) construction and their phylogenetic distributions were similar to those of sequences from DGGE. Approximately 80% of clones were affiliated to Proteobacteria, Bacteroidetes and Cyanobacteria. The possible influences of dynamic pollution and hydrological conditions in the Victoria Harbor area on the particle-attached and free-living bacterial community structures were discussed.

Establishment of a micro-particle bombardment transformation system for Dunaliella saliva

In this study, we chronicle the establishment of a novel transformation system for the unicellular marine green alga, Dunaliella salina. We introduced the CaMV35S promoter-GUS construct into D. saliva with a PDS1000/He micro-particle bombardment system. Forty eight h after transformation, via histochemical staining, we observed the transient expression of GUS in D. salina cells which had been bombarded under rupture-disc pressures of 450 psi and 900 psi. We observed no GUS activity in either the negative or the blank controls. Our findings indicated that the micro-particle bombardment method constituted a feasible approach to the genetic transformation of D. salina. We also conducted tests of the cells' sensitivity to seven antibiotics and one herbicide, and our results suggested that 20 mu g/ ml of Basta could inhibit cell growth completely. The bar gene, which encodes for phosphinothricin acetyltransferase and confers herbicide tolerance, was introduced into the cells via the above established method. The results of PCR and PCR-Southern blot analyses indicated that the gene was successfully integrated into the genome of the transformants.

Transient expression of lacZ in particle bombarded Gracilaria changii (Gracilariales, Rhodophyta)

Using a Biolistic PDS 1000/He system, healthy thalli of Gracilaria changii were bombarded with gold particles coated with plasmid DNA containing the lacZ reporter gene. Transient expression of lacZ was observed in bombarded thalli under the rupture-disc pressures of 4482, 6206, 7584 and 8963 KPa, two days after bombardment. Although G. changii exhibits a slight blue background, positive expression and the background colour can be clearly differentiated. The results indicate that lacZ could be a useful reporter gene and that SV40 promoter could be an effective promoter for Gracilaria transformation.

Genetic transformation in Kappaphycus alvarezii using micro-particle bombardment: a potential strategy for germplasm improvement

This study investigated the delivery of a SV40 promoter driving lacZ gene into cells of Kappaphycus alvarezii using particle bombardment. Thallus pieces 0.5-0.8 mm in diameter and 1 cm in length were prepared as gene recipients. Bombardment parameters of 450 psi (rupture pressures) x 6 cm (particle travel distances), 650 psi x 6 cm, 1,100 psi x 6 cm and 1,100 psi x 9 cm were used. A significant increase in transformation efficiency from about 33% under the rupture pressure of 450 psi to 87% at 650 psi was observed in transformed thalli. Most of the positive cells appeared in epidermal cells bombarded at 450 psi, whereas positive signals were seen in both epidermal and medullary cells at 650 psi. No positive transient expression was detected at a bombardment of 1,100 psi, or in negative or blank controls. For the conditions tested, the best parameter was obtained at 650 psi at a distance of 6 cm. Thus, the strategy of taking vegetative thalli as recipients, using particle bombardment, and combining this with micro-propagation, together with developing an in vivo selectable marker, is a viable way to produce stable transformants, to eliminate chimeric expression, and to achieve transgenic breeding in K. alvarezii.

Modification to the Hardisty equation, regarding the relationship between sediment transport rate and particle size

Bagnold-type bed-load equations are widely used for the determination of sediment transport rate in marine environments. The accuracy of these equations depends upon the definition of the coefficient k(1) in the equations, which is a function of particle size. Hardisty (1983) has attempted to establish the relationship between k(1) and particle size, but there is an error in his analytical result. Our reanalysis of the original flume data results in new formulae for the coefficient. Furthermore, we found that the k(1) values should be derived using u(1) and u(1cr) data; the use of the vertical mean velocity in flumes to replace u(1) will lead to considerably higher k(1) values and overestimation of sediment transport rates.

Simulating a typhoon storm surge in the East Sea of China using a coupled model

A coupled numerical model with a 2' x 2' resolution grid has been developed and used to simulate five typical typhoon storm surges (5612, 7413, 7910, 8114, and 9711) in the East Sea of China. Three main driving forces have been considered in this coupled model: wave radiation stress, combined wave-current bottom shear stress and wave-state-dependent surface wind stress. This model has then been compared with in situ measurements of the storm set-up. The effect of different driving force components on the total storm surge has also been investigated. This study has found that the coupled model with high resolution is capable of simulating the five typical typhoons better than the uncoupled models, and that the wave-dependent surface wind stress plays an important role in typhoon storm surge-wave coupling in this area and can increase the storm set-up by 1 m. The study of the five typhoon cases has shown that the general coupling effects could increase storm set-up by 20-32%. Thus, it is suggested that to predict typhoon storm surges in the East Sea of China, a storm surge-wave coupled model be adopted. (C) 2008 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.

An analysis of the recent severe storm surge disaster events in China

The economic loss caused by the storm surge disasters is much higher than that caused by any other marine disaster in China, the loss from the severe storm surge disaster being the highest. Statistics show that there were 62 typhoon landings over the east-southeast coast of China since 1990, three of which, occurring in 1992, 1994 and 1997, respectively, caused the most severe damage. The direct economic losses due to these events are 9.3, 17.0 and 30 billion yuan (RMB, or about 1.7, 2.6 and 3.8 billion USD, respectively), which is much greater than the loss of 5.5 billion yuan (RMB) on an average every year during the 1989-1991 period. This paper makes a comparative analysis of the damage caused by the three events and presents an overview of progress of precautions against storm surge disaster in China. The suggested counter measures to mitigate the loss from the severe storm surge disasters in China is as follows: (1) Raise the whole society awareness of precaution against severe storm surge disaster; (2) Work out a new plan for building sea walls; (3) Improve and perfect the available warning and disaster relief command system; (4) Develop the insurance service in order to promptly mitigate the loss caused by severe storm surge disaster event.

Effects of winds, tides and storm surges on ocean surface waves in the Sea of Japan

Ocean surface waves are strongly forced by high wind conditions associated with winter storms in the Sea of Japan. They are also modulated by tides and storm surges. The effects of the variability in surface wind forcing, tides and storm surges on the waves are investigated using a wave model, a high-resolution atmospheric mesoscale model and a hydrodynamic ocean circulation model. Five month-long wave model simulations are inducted to examine the sensitivity of ocean waves to various wind forcing fields, tides and storm surges during January 1997. Compared with observed mean wave parameters, results indicate that the high frequency variability in the surface wind filed has very great effect on wave simulation. Tides and storm surges have a significant impact on the waves in nearshores of the Tsushima-kaihyo, but not for other regions in the Sea of Japan. High spatial and temporal resolution and good quality surface wind products will be crucial for the prediction of surface waves in the JES and other marginal seas, especially near the coastal regions.

空间电流体系地磁反演及理论研究

The space currents definitely take effects on electromagnetic environment and also are scientific highlight in the space research. Space currents as a momentum and energy provider to Geospace Storm, disturb the varied part of geomagnetic field, distort magnetospheric configuration and furthermore take control of the coupling between magnetosphere and ionosphere. Due to both academic and commercial objectives above, we carry on geomagnetic inverse and theoretical studies about the space currents by using geomagnetic data from INTERMAGNET. At first, we apply a method of Natural Orthogonal Components (NOC) to decomposition the solar daily variation, especially for (solar quiet variation). NOC is just one of eign mode analysis, the most advantage of this method is that the basic functions (BFs) were not previously designated, but naturally came from the original data so that there are several BFs usually corresponding to the process really happened and have more physical meaning than the traditional spectrum analysis with the fixed BFs like Fourier trigonometric functions. The first two eign modes are corresponding to the and daily variation and their amplitudes both have the seasonal and day-to-day trend, that will be useful for evaluating geomagnetic activity indices. Because of the too strict constraints of orthogonality, we try to extend orthogonal contraints to the non-orthogonal ones in order to give more suitable and appropriate decomposition of the real processes when the most components did not satisfy orthogonality. We introduce a mapping matrix which can transform the real physical space to a new mathematical space, after that process, the modified components which associated with the physical processes have satisfied the orthogonality in the new mathematical space, furthermore, we can continue to use the NOC decomposition in the new mathematical space, and then all the components inversely transform back to original physical space, so that we would have finished the non-orthogonal decomposition which more generally in the real world. Secondly, geomagnetic inverse of the ring current’s topology is conducted. Configurational changes of the ring current in the magnetosphere lead to different patterns of disturbed ground field, so that the global configuration of ring current can be inferred from its geomagnetic perturbations. We took advantages of worldwide geomagnetic observatories network to investigate the disturbed geomagnetic field which produced by ring current. It was found that the ring current was not always centered at geomagnetic equator, and significantly deviated off the equator during several intense magnetic storms. The deviation owing to the tilting and latitudinal shifting of the ring current with respect to the earth’s dipole can be estimated from global geomagnetic survey. Furthermore those two configurational factors which gave a quantitative description of the ring current configuration, will be helpful to improve the Dst calibration and understand the dependence of ring current’s configuration on the plasma sheet location relative to the equator when magnetotail field warped. Thirdly, the energization and physical acceleration process of ring current during magnetic storm has been proposed. When IMF Bz component increase, the enhanced convection electric field drive the plasma injection into the inner magnetosphere. During the transport process, a dynamic heating is happened which make the particles more ‘hot’ when the injection is more deeply inward. The energy gradient along the injection path is equivalent to a kind of force, which resist the plasma more earthward injection, as a diamagnetic effect of the magnetosphere anti and repellent action to the exotically injected plasma. The acceleration efficiency has a power law form. We use analytical way to quantitatively describe the dynamical process by introducing a physical parameter: energization index, which will be useful to understand how the particle is heated. At the end, we give a scheme of how to get the from storm time geomagnetic data. During intense magnetic storms, the lognormal trend of geomagnetic Dst decreases depend on the heating dynamic of magnetosphere controlling ring current. The descending pattern of main phase is governed by the magnetospheric configuration, which can be describled by the energization index. The amplitude of Dst correlated with convection electric field or south component of the solar wind. Finally, the Dst index is predicted by upstream solar wind parameter. As we known space weather have posed many chanllenges and impacts on techinal system, the geomagnetic index for evaluating the activity space weather. We review the most popular Dst prediction method and repeat the Dst forecasting model works. A concise and convnient Key Points model of the polar region is also introduced to space weather. In summary, this paper contains some new quantitative and physical description of the space currents with special focus on the ring current. Whatever we do is just to gain a better understanding of the natural world, particularly the space environment around Earth through analytical deduction, algorithm designing and physical analysis, to quantitative interpretation. Applications of theoretical physics in conjunction with data analysis help us to understand the basic physical process govering the universe.

磁层-电离层耦合现象的分析研究

The magnetosphere-ionosphere coupling is mainly manifested by the trans- porting processes of energy into the ionosphere , the energy is carried by solar wind and firstly accumulate at the magnetosphere, and the coupling processes also significantly include the interaction between the magnetosphere and ionosphere for mass and energy. At the quiet condition, energy is delivered by the large-scale convection of the geomagnetic field; the huge energy from solar wind bulk will be injected into and consumed at the near magnetosphere and ionosphere by the geomagnetic storm and substorm activities. Aurorae and FACs (Field-aligned currents) are the important phenomena in the coupling processes. In the present work, firstly, we analyze the activity characteristics of auroral precipitating particle, secondly, we study the distribution characters of large-scale field aligned currents (LS FACs) at storm-time using the observations from different satellites at different altitudes. Finally, we investigate the evolution of the geomagnetic field configuration at the nightside sector on the onset of the expansion phase in a substorm event, the substorm event happened at 0430UT to 0630UT on 8th Nov. 2004. The main results as follows： At the first, the data of the estimated power input (EPI) of auroral particles from NOAA/POES (Polar orbiting environmental satellite) for some 30 years have been analyzed. The variation tendencies of the EPI generally coincide with aa, AE and Dst indices. The annual variation of EPI shows equinox peaks and an asymmetric-activity with a higher peak in the winter-hemisphere than in the summer-hemisphere. The diurnal UT variations are different from north and south hemisphere: for north hemisphere, the peak appears at 1200UT, and the relative deviation is 22% to the daily average of the north hemisphere. For south hemisphere, the maximal deviation is 22% at 2000UT. So the diurnal variation of EPI is more dominant than the annual variation which maximal deviation is 3% to 12% for different seasons. Studies on correlations of the hourly average of EPI, Pa, with AE and Dst indices show a correlation coefficient r=0.74 of Pa and AE, and r=-0.55 of Pa and Dst. The hourly EPIs for north and south polar regions, NPa and SPa, show a north-south asymmetry with a higher correlation of SPa and AE (or Dst). Time delays of EPI with respect to magnetic indices are examined, the maximum correlation coefficient of Pa with AE (r=0.78) occurs when the time delay =0, suggesting a synchronous activity of auroral electrojet and auroral precipitating particles, while =1-2h, the correlation coefficient of Pa with Dst is maximum (r=0.57), suggesting that the activity of auroral particle precipitating may influence the ring current on some extent. Sencondly, we use the high-resolution magnetic field vector data of the CHAMP satellite to investigate the distribution of large-scale FACs during the great magnetic storm on 7th to 8th Nov. 2004. The results show that, whether in the northern or southern hemisphere, the number and density of large-scale FACs during the main-phase are more and bigger than these during the recover-phase, and the number of large-scale FACs in morning sector obviously is more than that in afternoon sector. In terms of the magnetic indices, we find that large-scale FACs in morning sector significantly affected by the substorm activities, while in afternoon sector the large-scale FACs mainly indicate the fluctuations of the ring-current in storm time. Accordingly to the former studies, similarly, we find that in the morning sector, the scale of the large-scale FACs move to the high-latitude region, and in the afternoon sector, large-scale FACs distinctly expand to the low-latitude region. During the time periods that the NOAA/POES auroral precipitating particle power data temporally correspond to the large-scale FACs, the more the power of auroral particle is, the more and bigger the number and density of FACs are. At the same time, we use the magnetic field vector data of POLAR obtain a good form of region 1, region 2, and three pieces of cusp FACs during a single transit at 1930UT-2006UT on 07th. And the characteristics of simultaneous electric field and energy particles observations on Polar are coincide with the five FACs pieces. Finally, by means of the observation of Cluster 4 and Goes 10、 Goes 12, we analyze the evolution process of the change of the magnetic field configuration at night sector at the expansion phase of a substorm event which happened during 0430UT to 0630UT on 8th Nov. 2004, we find that the times of the beginning of the polarizations of magnetic field are observed from Goes 10 to Goes 12 then to Cluster 4. So, at the synchronous orbit ( 6.6 RE) to 10RE distance scale of the neutral sheet, the current disruption spread tailward. Simultaneously, the strengthen of the FACs deduced from these satellites’ magnetic field observations are almost consistent with the times of polarizations, as well as the high energy particles injection and the electric field dominant variation. The onset times determined by the magnetic field polarizations from these satellites are all ahead of the onset time that confirmed from the auroral electrojet indices. So, these characters of different observations can be used as the criterions to determine the onset time for the substorms of such type as we studied.