Expression of muscle-related genes and two MyoD genes during amphioxus notochord development

The notochord is one of the diagnostic features of the phylum Chordata. Despite the similarities in the early morphogenetic patterns of the notochords of various chordates, they are strikingly distinct from one another at the histological level. The amphioxus notochord is one example of an evolutionary novelty because it is made up of muscle cells. Our previous expressed sequence tag analysis, targeting messenger RNAs expressed in the adult amphioxus notochord, demonstrated that many muscle-related genes are expressed there. To characterize amphioxus notochord cells and to gain insights into the myogenic program in the notochord, we determined the spatial and temporal expression patterns of these muscle-related genes during amphioxus development. We found that BbNA1 (notochord actin), Amphi-Trop I (troponin I), Amphi-TPmyosin (tropomyosin), Amphi-MHC2 (myosin heavy chain), Amphi-nMRLC (notochord-specific myosin regulatory light chain), AmphinTitin/MLCK (notochord-specific titin/myosin light chain kinase), Amphi-MLP/CRP3 (muscle LIM protein), and Amphi-nCalponin (notochord-specific calponin) are expressed with characteristic patterns in notochord cells, including the central cells, dorsally located cells, and ventrally located cells, suggesting that each notochord cell has a unique molecular architecture that may reflect its function. In addition, we characterized two MyoD genes (Amphi-MyoD1 and Amphi-MyoD2) to gain insight into the genetic circuitry governing the formation of the notochord muscle. One of the MyoD genes (Amphi-MyoD2) is expressed in the central notochord cells, and the coexistence of Amphi-MyoD2 transcripts along with the Amphi-MLP/CRP3 transcripts implies the participation of Amphi-MyoD2 in the myogenic program in the notochord muscle.

Excitation energy distribution between two photosystems in Porphyra yezoensis and its significance in photosynthesis evolution

Comparative investigation on energy distribution between two photosystems were carried out in the sporophytes and gametophytes of Porphyra yezoensis. By performing 77 K fluorescence spectra, we suggested that there probably existed a pathway for energy transfer from PS II to PS I to redistribute the absorbed energy in gametophytes, while no such a way or at minor level in sporophytes. Electron transfer inhibitor DCMU blocked the energy transfer from PS II to PS I in gametophytes, but no obvious effects on sporophytes. These indicated that excitation energy distribution between two photosystems in gametophytes was more cooperative than that in sporophytes. These data in ontogenesis reflected the evolution process of photosynthetic organisms and supported the hypothesis of independent evolution of each photosystem.

There are two 5 '-flanking regions of bkt encoding beta-carotene ketolase in Haematococcus pluvialis

The unicellular green alga Haematococcus pluvialis accumulates a commercially valuable astaxanthin, with levels reaching up to 4% dry weight under environmental stress. In recent years, much effort has been devoted to understanding the molecular mechanisms regulating astaxanthin biosynthetic pathways. Beta-carotene ketolase (bkt), with control being exhibited at the transcription level, plays an important role in astaxanthin biosynthesis by H. pluvialis. Here we demonstrate the presence of two separate 5'-flanking regions [1.5 kilobase (kb) and 2 kb] of bkt (bkt1 and bkt2) that possess regulatory elements similar to those of known stress-responsive genes in plants. Results of 5'-deletion constructs and transient beta-galactosidase expression assays demonstrate that there may be positive regulatory elements governing expression in the shorter promoter at -1060/-900 from the 1.5 kb 5' region, and in the longer promoter at -1838/-1219 and at -1046/ -734 from the 2 kb 5' region relative to each homologous ATG start codon. Furthermore, our present studies reveal that the first intron (+371/+497) downstream from the 1.5 kb 5' untranslated region of bkt1 may function as a negative regulatory element to regulate its own promoter.

Comparative growth and viability of hybrids between two populations of Chinese shrimp (Fennropenaeus chinensis)

Fenneropenaeus chinensis is confined to the Yellow Sea and Bohai Sea in China and the West Coast of the Korean Peninsula. Intra- and intercross populations were produced between Rushany (YP) and Korean (KN) populations. Seven traits were recorded. The heterosis of hybrids was computed and comparison between treatments was performed by ANOVA. At the fourth month after post-larvae, six indexes of growth trait and viability showed a range of heterosis, ranging from 0.514% to 14.950%. At the fifth month after post-larvae, six indexes of growth trait and viability ranged from -9.000% to 19.090%. The negative heterosis was observed in CL, HST and viability. The heterosis of KN female xYP male tended generally to increase as the age of the Chinese shrimp increased while the heterosis of YP female xKN male tended to decrease. The results indicated that the viability of reciprocal hybrids were not significantly different (P > 0.05) from their parents during the experiment. The result of ANOVA indicated that the F1 hybrids were significantly different (P < 0.05) in WST and TW at the fourth month. The multiple comparisons of LSD test indicated that the hybrids of YP female xKN male were significantly different (P < 0.05) from their parents in TW. The hybrids of YP female xKN male were significantly different (P < 0.05) from the other three combinations in WST. At the fifth months, the F1 hybrids had significant difference (P < 0.05) in body weight while other traits showed no significant differences (P > 0.05) from their parents. The multiple comparisons of LSD test indicated that the hybrids of KN female xYP male were significantly different (P < 0.05) from the KN parents in TW. The results indicate that in experimental conditions, the F-1 hybrids created from two populations of Chinese shrimp showed a certain level of heterosis for growth performance and viability. The crossing scheme may improve growth performance and viability in Chinese shrimp, but the improvement may be limited because effective crossbreeding requires the maintenance of pure, preferably inbred, lines and possibly involves specialized sire and dam lines. Therefore, the exploitation of heterosis through single crossbreeding in Chinese shrimp is of limited utility in practical commercial shrimp aquaculture in spite of the potential of significant heterosis. The crossbreeding of different populations can be applied in the establishment of base populations.

Dynamics of intraseasonal sea level and thermocline variability in the equatorial Atlantic during 2002-03

Satellite and in situ observations in the equatorial Atlantic Ocean during 2002-03 show dominant spectral peaks at 40-60 days and secondary peaks at 10-40 days in sea level and thermocline within the intraseasonal period band (10-80 days). A detailed investigation of the dynamics of the intraseasonal variations is carried out using an ocean general circulation model, namely, the Hybrid Coordinate Ocean Model (HYCOM). Two parallel experiments are performed in the tropical Atlantic Ocean basin for the period 2000-03: one is forced by daily scatterometer winds from the Quick Scatterometer (QuikSCAT) satellite together with other forcing fields, and the other is forced by the low-passed 80-day version of the above fields. To help in understanding the role played by the wind-driven equatorial waves, a linear continuously stratified ocean model is also used. Within 3 degrees S-3 degrees N of the equatorial region, the strong 40-60-day sea surface height anomaly (SSHA) and thermocline variability result mainly from the first and second baroclinic modes equatorial Kelvin waves that are forced by intraseasonal zonal winds, with the second baroclinic mode playing a more important role. Sharp 40-50-day peaks of zonal and meridional winds appear in both the QuikSCAT and Pilot Research Moored Array in the Tropical Atlantic (PIRATA) data for the period 2002-03, and they are especially strong in 2002. Zonal wind anomaly in the central-western equatorial basin for the period 2000-06 is significantly correlated with SSHA across the equatorial basin, with simultaneous/ lag correlation ranging from-0.62 to 0.74 above 95% significance. Away from the equator (3 degrees-5 degrees N), however, sea level and thermocline variations in the 40-60-day band are caused largely by tropical instability waves (TIWs). On 10-40-day time scales and west of 10 degrees W, the spectral power of sea level and thermocline appears to be dominated by TIWs within 5 degrees S-5 degrees N of the equatorial region. The wind-driven circulation, however, also provides a significant contribution. Interestingly, east of 10 W, SSHA and thermocline variations at 10 40- day periods result almost entirely from wind-driven equatorial waves. During the boreal spring of 2002 when TIWs are weak, Kelvin waves dominate the SSHA across the equatorial basin (2 degrees S-2 degrees N). The observed quasi-biweekly Yanai waves are excited mainly by the quasi-biweekly meridional winds, and they contribute significantly to the SSHA and thermocline variations in 1 degrees-5 degrees N and 1 degrees-5 degrees S regions.

Long-Wave Dynamics of Sea Level Variations during Indian Ocean Dipole Events

Long-wave dynamics of the interannual variations of the equatorial Indian Ocean circulation are studied using an ocean general circulation model forced by the assimilated surface winds and heat flux of the European Centre for Medium-Range Weather Forecasts. The simulation has reproduced the sea level anomalies of the Ocean Topography Experiment (TOPEX)/Poseidon altimeter observations well. The equatorial Kelvin and Rossby waves decomposed from the model simulation show that western boundary reflections provide important negative feedbacks to the evolution of the upwelling currents off the Java coast during Indian Ocean dipole (IOD) events. Two downwelling Kelvin wave pulses are generated at the western boundary during IOD events: the first is reflected from the equatorial Rossby waves and the second from the off-equatorial Rossby waves in the southern Indian Ocean. The upwelling in the eastern basin during the 1997-98 IOD event is weakened by the first Kelvin wave pulse and terminated by the second. In comparison, the upwelling during the 1994 IOD event is terminated by the first Kelvin wave pulse because the southeasterly winds off the Java coast are weak at the end of 1994. The atmospheric intraseasonal forcing, which plays an important role in inducing Java upwelling during the early stage of an IOD event, is found to play a minor role in terminating the upwelling off the Java coast because the intraseasonal winds are either weak or absent during the IOD mature phase. The equatorial wave analyses suggest that the upwelling off the Java coast during IOD events is terminated primarily by western boundary reflections.

COASTAL SEA-LEVEL AND THE LARGE-SCALE CLIMATE STATE - A DOWNSCALING EXERCISE FOR THE JAPANESE ISLANDS

A major problem which is envisaged in the course of man-made climate change is sea-level rise. The global aspect of the thermal expansion of the sea water likely is reasonably well simulated by present day climate models; the variation of sea level, due to variations of the regional atmospheric forcing and of the large-scale oceanic circulation, is not adequately simulated by a global climate model because of insufficient spatial resolution. A method to infer the coastal aspects of sea level change is to use a statistical ''downscaling'' strategy: a linear statistical model is built upon a multi-year data set of local sea level data and of large-scale oceanic and/or atmospheric data such as sea-surface temperature or sea-level air-pressure. We apply this idea to sea level along the Japanese coast. The sea level is related to regional and North Pacific sea-surface temperature and sea-level air pressure. Two relevant processes are identified. One process is the local wind set-up of water due to regional low-frequency wind anomalies; the other is a planetary scale atmosphere-ocean interaction which takes place in the eastern North Pacific.

潮波模型的优化开边界方法研究

数值模式是潮波研究的一种有利手段，但在研究中会面临各种具体问题，包括开边界条件的确定、底摩擦系数和耗散系数的选取等。数据同化是解决这些问题的一种途径，即利用有限数量的潮汐观测资料对潮波进行最优估计，其根本目的是迫使模型预报值逼近观测值，使模式不要偏离实际情况太远。本文采用了一种优化开边界方法，沿着数值模型的开边界优化潮汐水位信息，目的是设法使数值解在动力约束的意义下接近观测值，获得研究区域的潮汐结果。边界值由指定优化问题的解来定，以提高模拟区域的潮汐精度，最优问题的解是基于通过开边界的能量通量的变化，处理开边界处的观测值与计算值之差的最小化。这里提供了辐射型边界条件，由Reid 和Bodine（本文简称为RB）推导，我们将采用的优化后的RB方法（称为ORB）是优化开边界的特殊情况。 本文对理想矩形海域（ E- E， N- N， 分辨率 ）进行了潮波模拟，有东部开边界，模式采用ECOM3D模式。对数据结果的误差分析采用，振幅平均偏差，平均绝对偏差，平均相对误差和均方根偏差四个值来衡量模拟结果的好坏程度。 需要优化入开边界的解析潮汐值本文采用的解析解由方国洪《海湾的潮汐与潮流》（1966年）方法提供，为验证本文所做的解析解和方文的一致，本文做了其第一个例子的关键值a,b,z,结果与其结果吻合的相当好。但略有差别，分析的可能原因是两法在具体迭代方案和计算机保留小数上有区别造成微小误差。另外，我们取m=20，得到更精确的数值，我们发现对前十项的各项参数值，取m=10,m=20各项参数略有改进。当然我们可以获得m更大的各项参数值。 同时为了检验解析解的正确性讨论m和l变化对边界值的影响，结果指出，增大m，m=20时，u的模最大在本身u1或u2的模的6%；m=100时，u的模最大在本身u1或u2的模的4%；m再增大，m=1000时，u的模最大在本身u1或u2的模的4%，改变不大。当l<1时， =0处u的模最大为2。当l=1时， =0处u的模最大为0.1，当l>1时，l越大，u的模越小，当l=10时，u的模最大为0.001，可以认为为0。 为检验该优化方法的应用情况，我们对理想矩形区域进行模拟，首先将本文所采用的优化开边界方法应用于30m的情况，在开边界优化入开边界得出模式解，所得模拟结果与解析解吻合得相当好，该模式解和解析解在整个区域上，振幅平均绝对偏差为9.9cm,相位平均绝对偏差只有4.0 ，均方根偏差只有13.3cm，说明该优化方法在潮波模型中有效。 为验证该优化方法在各种条件下的模拟结果情况，在下面我们做了三类敏感性试验： 第一类试验：为证明在开边界上使用优化方法相比于没有采用优化方法的模拟解更接近于解析解，我们来比较ORB条件与RB条件的优劣，我们模拟用了两个不同的摩擦系数，k分别为：0,0.00006。 结果显示，针对不同摩擦系数，显示在开边界上使用ORB条件的解比使用RB条件的解无论是振幅还是相位都有显著改善，两个试验均方根偏差优化程度分别为84.3%，83.7%。说明在开边界上使用优化方法相比于没有采用优化方法的模拟解更接近于解析解，大大提高了模拟水平。上述的两个试验得出， k=0.00006优化结果比k=0的好。 第二类试验，使用ORB条件确定优化开边界情况下，在东西边界加入出入流的情况，流考虑线性和非线性情况，结果显示，加入流的情况，潮汐模拟的效果降低不少，流为1Sv的情况要比5Sv的情况均方根偏差相差20cm,而不加流的情况只有0.2cm。线性流和非线性流情况两者模式解相差不大，振幅，相位各项指数都相近, 说明流的线性与否对结果影响不大。 第三类试验，不仅在开边界使用ORB条件，在模式内部也使用ORB条件，比较了内部优化和不优化情况与解析解的偏差。结果显示，选用不同的k,振幅都能得到很好的模拟，而相位相对较差。另外，在内部优化的情况下，考虑不同的k的模式解， 我们选用了与解析解相近的6个模式解的k,结果显示，不同的k,振幅都能得到很好的模拟，而相位较差。 总之，在开边界使用ORB条件比使用RB条件好，振幅相位都有大幅度改进，在加入出入流情况下，流的大小对模拟结果有影响，但线形流和非线性流差别不大。内部优化的结果显示，模式采用不同的k都能很好模拟解析解的振幅。

Estuarine nutrient loading affects phytoplankton growth and microzooplankton grazing at two contrasting sites in Hong Kong coastal waters

To investigate the effects of enhanced nutrient loading in estuarine waters on phytoplankton growth and microzooplankton grazing, we conducted monthly dilution experiments at 2 stations in Hong Kong coastal waters with contrasting trophic conditions. The western estuarine station (WE) near the Pearl River estuary is strongly influenced by freshwater discharge, while the eastern oceanic station (EO) is mostly affected by the South China Sea. Growth rates of phytoplankton were often limited by nutrients at EO, while nutrient limitation of phytoplankton growth seldom Occurred at WE due to the high level of nutrients delivered by the Pearl River, especially in the summer rainy season. Higher chlorophyll a, microzooplankton biomass, phytoplankton growth and microzooplankton grazing rates were found at WE than at EO. However, the increase in chlorophyll greatly exceeded the increase in phytoplankton growth rate, reflecting different response relationships to nutrient availability. Strong seasonality was observed at both stations, with temperature being an important factor affecting both phytoplankton growth and microzooplankton grazing rates. Picophytoplankton, especially Synechococcus, also exhibited great seasonality at EO, with summer abundances being 2 or 3 orders of magnitude higher than those during winter, Our results confirm that in eutrophic coastal environments, microzooplankton grazing is a dominant loss pathway for phytoplankton, accounting for the utilization of >50%, of primary production on average.

Photosynthesis-irradiance response at physiological level: A mechanistic model

A mechanistic model is developed to present the photosynthetic response of phytoplankton to irradiance at the physiological level. The model is operated on photosynthetic units (PSU), and each PSU is assumed to have two states: reactive and activated. Light absorption that drives a reactive PSU into the activated state results from the effective absorption of the PSU. Transitions between the two states are asymmetrical in rate. A PSU in the reactive state becomes activated much faster than it recovers from the activated state to the reactive one. The turnover time for an activated PSU to transit into the reactive one is defined by the turnover time of the electron transport chain. The present model yields a photosynthesis-irradiance curve (PE-curve) in a hyperbola, which is described by three physiological parameters: effective cross-section (sigma (PSII)), turnover time of electron transport chain (tau) and number of PSUs (N). The PE-curve has an initial slope of sigma (PSII) x N, a half-saturated irradiance of 1/(sigma (PSII)), and a maximal photosynthetic rate of Nlc at the saturated irradiance. The PE-curve from the present model is comparable to the empirical function based on the target theory described by the Poisson distribution. (C) 2001 Academic Press.

Food sources of three bivalves living in two habitats of Jiaozhou Bay (Qingdao, China): Indicated by lipid biomarkers and stable isotope analysis

Food Sources of three filter-feeding bivalves from two habitats (intertidal oyster Crassostrea gigas, mussel Mytilus galloprovincialis. and subtidal cultured scallop Chlamys farreri) of Jiaozhou Bay (Qingdao,China) were determined by fatty acid and stable isotope in analysis. Cultured scallop was characterized by significant diatom markets such as 16:1/16:0 close to 1 and high ratio of 20:5(n - 3)/22:6(n - 3), hence we assume that the scallop mainly feeds on diatoms. Fatty acid biomarkers specific to bacteria and terrestrial materials were also found in considerable amounts in scallop tissue, which suggested that there were Substantial bacterial and terrestrial input into the food of the species. Intertidal oyster and mussel, however, exhibited significant flagellate marker. 22:6(n - 3). and lower level of diatom markers. which indicated that flagellates are also part of intertidal bivalves' Planktonic food Sources: meanwhile, high level of Chlorophyta fatty acid marker, Sigma 18:2(n - 6) + 18:3(n - 3), suggested that Ulva pertusa (Chlorophyta) seaweed bed supplied important food sources to intertidal bivalves. Additionally, result of stable isotope analysis showed that phytoplankton contributed 86.2 to 89.0% to intertidal bivalves' carbon budget; macroalga U. pertusa origin source had a contribution of MIX, to 11.0%, which indicated its role Lis in important supplemental food source to intertidal bivalves. From this study. it is concluded that the dietary difference of three bivalves probably relates to the different potential food sources in the scallop farm and intertidal zone in Jiaozhou Bay.

Inbreeding depression for various traits in two cultured populations of the American bay scallop, Argopecten irradians irradians Lamarck (1819) introduced into China

This paper examines the effect of inbreeding level of population on the magnitude of inbreeding depression expressed by comparing them between two cultured populations (A and B) in the hermaphroditic animal of the bay scallop Argopecten irradians irradians. Population A is expected to have less genetic variations and higher inbreeding level due to longer cultured history (20 generations) and less "ancestral" individuals (26 individuals) than population B due to shorter cultured history (4 generations) and more "ancestral" individuals (406 individuals). Two groups within each population were produced, one using self-fertilization and one using mass-mating within the same population. Selfed offspring (AS and BS) from two populations both had lower fitness components than their mass-mated counterparts (AM and BM) and exhibited inbreeding depression for all examined traits, e.g. lower hatching, less viability and slower growth, indicating that inbreeding depression is a common feature in this animal. Fitness components in all traits of offspring from population A significantly differed those from population B and the magnitude of inbreeding depression for all traits in population A with higher inbreeding level was significantly smaller than that in population B with lower inbreeding level, indicating that both fitness components and magnitude of inbreeding depression were significantly affected by inbreeding level of populations and genetic load harbored in population A may be partially purged through inbreeding. Moreover, the magnitude of inbreeding depression in the two populations both varied among traits and life history stages. The present results support the partial-dominance hypothesis of inbreeding depression. (C) 2008 Elsevier B.V. All rights reserved.

接收函数方法与中国及邻区上地幔间断面的研究

The receiver function method applied in researching the discontinuities in upper mantle was systematically studied in this paper. Using the theoretical receiver functions, the characteristics of P410S and P660S phases were analyzed, and the influencing factors for detection of these phases were discussed. The stability of receiver function was studied, and a new computational method of receiver function, RFSSMS (Receiver Function of Stack and Smooth of Multi seismic-records at a Single station), was put forward. We built initial reference velocity model for the media beneath each of 18 seismic stations respectively; then estimated the buried depths of 410-km and 660-km discontinuities(simply marked as '410' and '660') under the stations by using the arrive time differences of P410S and P660S with P. We developed a new receiver function inversion method -PGARFI (Peeling-Genetic Algorithm of Receiver Function Inversion), to obtain the whole crust and upper mantle velocity structure and the depths of discontinuities beneath a station. The major works and results could be summarized as follows: (1) By analysis of the theoretical receiver functions with different velocity models and different ray parameters, we obtain the knowledge: The amplitudes of P410S and P660S phases are decreasing with the increasing of epicentral distance A , and the arrival time differences of these phases with P are shorter as A is longer. The multiple refracted and/or reflected waves yielded on Moho and the discontinuities in the crust interfere the identification of P410S. If existing LVZ under the lithosphere, some multiple waves caused by LVZ will interfere the identification of P410S. The multiple waves produced by discontinuity lied near 120km depth will mix with P410s phase in some range of epicentral distance; and the multiple waves concerned with the discontinuity lied near 210km depth will interfere the identification of P660S. The epicentral distance for P4i0s identification is limited, the upper limit is 80° . The identification of P660S is not restricted by the epicenter distance obviously. The identification of P410S and P6gos in the theoretical receiver functions is interfered weakly from the seismic wave attenuation caused by the media absorption if the Q value in a reasonable range. (2) The stability of receiver function was studied by using synthetic seismograms with different kind of noise. The results show that on the condition of high signal-noise-ratio of seismic records, the high frequency background noise and the low frequency microseism noise do not influence the calculating result of receiver function. But the media "scattering noise" influence the stability of receiver function. When the scattering effect reach some level, the identification of P4iOs and P66os is difficult in single receiver function which is yielded from only one seismic record. We provided a new method to calculate receiver function, that is, with a group of earthquake records, stacking the R and Z components respectively in the frequency domain, and weighted smooth the stacked Z component, then compute the complex spectrum ratio of R to Z. This method can improve the stability of receiver function and protrude the P4i0s and P66os in the receiver function curves. (3) 263 receiver functions were provided from 1364 three component broadband seismograms recorded at 18 stations in China and adjacent areas for the tele-earthquakes. The observed arrival time differences of P410S and P660S with P were obtained in these receiver functions. The initial velocity model for every station was built according to the prior research results. The buried depths of '410' and '660' under a station were acquired by the way of adjusting the depths of these two discontinuities in the initial velocity model until the theoretical arrival time differences of P410S and P660S with P well conformed to the observed. The results show an obvious lateral heterogeneity of buried depths of ' 410' and (660' . The depth of '410' is shallower beneath BJI, XAN, LZH and ENH, but deeper under QIZ and CHTO, and the average is 403km . The average depth of '660' is 663km, deeper under MDJ and MAJO, but shallower under QIZ and HYB. (4) For inversing the whole crust and upper mantle velocity structure, a new inversion method -PGARFI (Peeling-Genetic Algorithm of Receiver Function Inversion) has- been developed here. The media beneath a station is divided into segments, then the velocity structure is inversed from receiver function from surface to deep successively. Using PGARFI, the multi reflection / refraction phases of shallower discontinuities are isolated from the first order refraction transform phase of deep discontinuity. The genetic algorithm with floating-point coding was used hi the inversion of every segment, and arithmetical crossover and non-uniform mutation technologies were employed in the genetic optimization. 10 independent inversions are completed for every segment, and 50 most excellent velocity models are selected according to the priority of fitness from all models produced in the inversion process. The final velocity structure of every segment is obtained from the weighted average of these 50 models. Before inversion, a wide range of velocity variation with depth and depth range of the main discontinuities are given according to priori knowledge. PGARFI was verified with numerical test and applied in the inversion of the velocity structure beneath HIA station down to 700km depth.

Total analytical method for Radix astragali extract using two-binary multi-segment gradient elution liquid chromatography

There is an urgent need for thorough analysis of Radix astragali, a widely used Chinese herb, for quality control purposes. This paper describes the development of a total analytical method for Radix astragali extract, a multi-component complex mixture. Twenty-four components were separated step by step from the extract using a series of isocratic isopropanol-methanol elutions, and then 42 components were separated similarly using methanol-water elutions. Based on the log k(w) and -S of the 66 components obtained from the above procedure and the optimization software developed in our laboratory, an optimum elution program consisting of seven methanol-water segments and four isopropanol-methanol segments was developed to finish the task of analyzing the total components in a single run. Under optimized gradient conditions, the sample of Radix astragali extract was analyzed. As expected, most of the components were well separated and the experimental chromatogram was in a good agreement with the predicted one.

A two-stage process with temperature-shift for enhanced anthocyanin production in strawberry cell suspension cultures

A two-stage process with temperature-shift has been developed to enhance the anthocyanin yield in suspension cultures of strawberry cells. The effect of the temperature-shift interval and the shift-time point was investigated for the optimization of this strategy. In this process, strawberry cells were grown at 30 degrees C (the optimum temperature for cell growth) for a certain period as the first stage, with the temperature shifted to a lower temperature for the second stage. In response to the temperature shift-down, anthocyanin synthesis was stimulated and a higher content could be achieved than that at both boundary temperatures but cell growth was suppressed. When the lower boundary temperature was decreased, cell growth was lowered and a delayed, sustained maximum anthocyanin content was achieved. Anthocyanin synthesis was strongly influenced by the shift-time point but cell growth was not. Consequently, the maximum anthocyanin content of 2.7 mg.g-fresh cell(-1) was obtained on day 9 by a temperature-shift from 30 degrees C, after 3-d culture, to 15 degrees C. The highest anthocyanin yield of 318 mg.L-1 on day 12 was achieved when the temperature was shifted from 30 degrees C, after 5-d culture, to 20 degrees C. For a global optimization of both the yield and productivity, the optimum anthocyanin yield and productivity of 272 mg.L-1 and 30.2 mg.L-1.d(-1) on day 9 were achieved by a two-stage culture with a temperature-shift from 30 degrees C after 3 d to 20 degrees C.