ANALYTICAL DEVICE FOR TEST FLUIDS, COMPRISES A CONDUCTIVE POLYMER COMPOSITION

A sensor for chemical species or biological species or radiation presenting to test fluid a polymer composition comprises polymer and conductive filler metal, alloy or reduced metal oxide and having a first level of electrical conductance when quiescent and being convertible to a second level of conductance by change of stress applied by stretching or compression or electric field, in which the polymer composition is characterised by at least one of the features in the form of particles at least 90% w/w held on a 100 mesh sieve; and/or comprising a permeable body extending across a channel of fluid flow; and/or affording in-and-out diffusion of test fluid and/or mechanically coupled to a workpiece of polymer swellable by a constituent of test fluid.

Velocity profiles in a cylindrical liquid jet by reconstructed velocimetry

An experimental setup and a simple reconstruction method are presented to measure velocity fields inside slightly tapering cylindrical liquid jets traveling through still air. Particle image velocimetry algorithms are used to calculate velocity fields from high speed images of jets of transparent liquid containing seed particles. An inner central plane is illuminated by a laser sheet pointed at the center of the jet and visualized through the jet by a high speed camera. Optical distortions produced by the shape of the jet and the difference between the refractive index of the fluid and the surrounding air are corrected by using a ray tracing method. The effect of the jet speed on the velocity fields is investigated at four jet speeds. The relaxation rate for the velocity profile downstream of the nozzle exit is reasonably consistent with theoretical expectations for the low Reynolds numbers and the fluid used, although the velocity profiles are considerably flatter than expected. © 2012 American Society of Mechanical Engineers.

A comparison of the value of viscosity for several water models using Poiseuille flow in a nano-channel.

The viscosity-temperature relation is determined for the water models SPC/E, TIP4P, TIP4P/Ew, and TIP4P/2005 by considering Poiseuille flow inside a nano-channel using molecular dynamics. The viscosity is determined by fitting the resulting velocity profile (away from the walls) to the continuum solution for a Newtonian fluid and then compared to experimental values. The results show that the TIP4P/2005 model gives the best prediction of the viscosity for the complete range of temperatures for liquid water, and thus it is the preferred water model of these considered here for simulations where the magnitude of viscosity is crucial. On the other hand, with the TIP4P model, the viscosity is severely underpredicted, and overall the model performed worst, whereas the SPC/E and TIP4P/Ew models perform moderately.

Thylakoid terminal oxidases are essential for the cyanobacterium Synechocystis sp. PCC 6803 to survive rapidly changing light intensities.

Cyanobacteria perform photosynthesis and respiration in the thylakoid membrane, suggesting that the two processes are interlinked. However, the role of the respiratory electron transfer chain under natural environmental conditions has not been established. Through targeted gene disruption, mutants of Synechocystis sp. PCC 6803 were generated that lacked combinations of the three terminal oxidases: the thylakoid membrane-localized cytochrome c oxidase (COX) and quinol oxidase (Cyd) and the cytoplasmic membrane-localized alternative respiratory terminal oxidase. All strains demonstrated similar growth under continuous moderate or high light or 12-h moderate-light/dark square-wave cycles. However, under 12-h high-light/dark square-wave cycles, the COX/Cyd mutant displayed impaired growth and was completely photobleached after approximately 2 d. In contrast, use of sinusoidal light/dark cycles to simulate natural diurnal conditions resulted in little photobleaching, although growth was slower. Under high-light/dark square-wave cycles, the COX/Cyd mutant suffered a significant loss of photosynthetic efficiency during dark periods, a greater level of oxidative stress, and reduced glycogen degradation compared with the wild type. The mutant was susceptible to photoinhibition under pulsing but not constant light. These findings confirm a role for thylakoid-localized terminal oxidases in efficient dark respiration, reduction of oxidative stress, and accommodation of sudden light changes, demonstrating the strong selective pressure to maintain linked photosynthetic and respiratory electron chains within the thylakoid membrane. To our knowledge, this study is the first to report a phenotypic difference in growth between terminal oxidase mutants and wild-type cells and highlights the need to examine mutant phenotypes under a range of conditions.

Winglets for Improved Aerothermal Performance of High Pressure Turbines

This paper investigates the design of winglet tips for unshrouded high pressure turbine rotors, considering aerodynamic and thermal performance simultaneously. A novel parameterization method has been developed to alter the tip geometry of a rotor blade. A design survey of un-cooled, flat-tipped winglets is performed using RANS calculations for a single rotor at engine representative operating conditions. Compared to a plain tip, large efficiency gains can be realized by employing an overhang around the full perimeter of the blade, but the overall heat load rises significantly. By employing an overhang on only the early suction surface, significant efficiency improvements can be obtained without increasing the overall heat transfer to the blade. The flow physics are explored in detail to explain the results. For a plain tip, the leakage and passage vortices interact to create a three-dimensional impingement onto the blade suction surface, causing high heat transfer. The addition of an overhang on the early suction surface displaces the tip leakage vortex away from the blade, weakening the impingement effect and reducing the heat transfer on the blade. The winglets reduce the aerodynamic losses by unloading the tip section, reducing the leakage flow rate, turning the leakage flow in a more streamwise direction and reducing the interaction between the leakage fluid and endwall flows. Generally these effects are most effective close to the leading edge of the tip, where the leakage flow is subsonic.

Application of methyl parathion hydrolase (MPH) as a labeling enzyme

Methyl parathion hydrolase (MPH) is an enzyme that catalyzes the degradation of methyl parathion, generating a yellow product with specific absorption at 405 nm. The application of MPH as a new labeling enzyme was illustrated in this study. The key advantages of using MPH as a labeling enzyme are as follows: (1) unlike alkaline phosphatase (AP), horseradish peroxidase (HRP), and glucose oxidase (GOD), MPH is rarely found in animal cells, and it therefore produces less background noise; (2) its active form in solution is the monomer, with a molecular weight of 37 kDa; (3) its turnover number is 114.70 +/- 13.19 s(-1), which is sufficiently high to yield a significant signal for sensitive detection; and (4) its 3D structure is known and its C-terminal that is exposed to the surface can be easily subjected to the construction of genetic engineering monocloning antibody-enzyme fusion for enzyme-linked immunosorbent assay (ELISA). To demonstrate its utility, MPH was ligated to an single-chain variable fragment (scFv), known as A1E, against a white spot syndrome virus (WSSV) with the insertion of a [-(Gly-Ser)(5)-] linker peptide. The resulting fusion protein MPH-A1E possessed both the binding specificity of the scFv segment and the catalytic activity of the MPH segment. When MPH-A1E was used as an ELISA reagent, 25 ng purified WSSV was detected; this was similar to the detection sensitivity obtained using A1E scFv and the HRP/Anti-E Tag Conjugate protocol. The fusion protein also recognized the WSSV in 1 mu L hemolymph from an infected shrimp and differentiated it from a healthy shrimp.

Initiation of immersed granular avalanches.

By means of coupled molecular dynamics-computational fluid dynamics simulations, we analyze the initiation of avalanches in a granular bed of spherical particles immersed in a viscous fluid and inclined above its angle of repose. In quantitative agreement with experiments, we find that the bed is unstable for a packing fraction below 0.59 but is stabilized above this packing fraction by negative excess pore pressure induced by the effect of dilatancy. From detailed numerical data, we explore the time evolution of shear strain, packing fraction, excess pore pressures, and granular microstructure in this creeplike pressure redistribution regime, and we show that they scale excellently with a characteristic time extracted from a model based on the balance of granular stresses in the presence of a negative excess pressure and its interplay with dilatancy. The cumulative shear strain at failure is found to be ≃ 0.2, in close agreement with the experiments, irrespective of the initial packing fraction and inclination angle. Remarkably, the avalanche is triggered when dilatancy vanishes instantly as a result of fluctuations while the average dilatancy is still positive (expanding bed) with a packing fraction that declines with the initial packing fraction. Another nontrivial feature of this creeplike regime is that, in contrast to dry granular materials, the internal friction angle of the bed at failure is independent of dilatancy but depends on the inclination angle, leading therefore to a nonlinear dependence of the excess pore pressure on the inclination angle. We show that this behavior may be described in terms of the contact network anisotropy, which increases with a nearly constant connectivity and levels off at a value (critical state) that increases with the inclination angle. These features suggest that the behavior of immersed granular materials is controlled not only directly by hydrodynamic forces acting on the particles but also by the influence of the fluid on the granular microstructure.

Study on methane hydration process in a semi-continuous stirred tank reactor

The methane hydration process is investigated in a semi-continuous stirred tank reactor. Liquid temperatures and reaction rates without stirrer are compared with those occurring with stirrer, while at the same time better stirring conditions of the methane hydration process are given by the experiments. Some basic data of fluid mechanics, for example, stirring Reynolds number, Froucle number and stirrer power, are calculated during the methane hydration process, which can be applied to evaluate stirrer capacity and provide some basic data for a scaled up reactor. Based on experiment and calculations in this work, some conclusions are drawn. First, the stirrer has great influence on the methane hydration process. Batch stirring is helpful to improve the mass transfer and heat transfer performances of the methane hydration process. Second, induction time can be shortened effectively by use of the stirrer. Third, in this paper, the appropriate stirring velocity and stirring time were 320 rpm and 30 min, respectively, at 5.0 MPa, for which the storage capacity and reaction time were 159.1 V/V and 370 min, respectively. Under the condition of the on-flow state, the initial stirring Reynolds number of the fluid and the stirring power were 12,150 and 0.54 W, respectively. Fourth, some suggestions, for example, the use of another type of stirrer or some baffles, are proposed to accelerate the methane hydration process. Comparing with literature data, higher storage capacity and hydration rate are achieved in this work. Moreover, some fluid mechanics parameters are calculated, which can provide some references to engineering application.

Hydroelastic dynamic characteristics of a slender axis-symmetric body

The slender axis-symmetric submarine body moving in the vertical plane is the object of our investigation. A coupling model is developed where displacements of a solid body as a Euler beam (consisting of rigid motions and elastic deformations) and fluid pressures are employed as basic independent variables, including the interaction between hydrodynamic forces and structure dynamic forces. Firstly the hydrodynamic forces, depending on and conversely influencing body motions, are taken into account as the governing equations. The expressions of fluid pressure are derived based on the potential theory. The characteristics of fluid pressure, including its components, distribution and effect on structure dynamics, are analyzed. Then the coupling model is solved numerically by means of a finite element method (FEM). This avoids the complicacy, combining CFD (fluid) and FEM (structure), of direct numerical simulation, and allows the body with a non-strict ideal shape so as to be more suitable for practical engineering. An illustrative example is given in which the hydroelastic dynamic characteristics, natural frequencies and modes of a submarine body are analyzed and compared with experimental results. Satisfactory agreement is observed and the model presented in this paper is shown to be valid.

花椒叶和土壤浸提液对土壤微生物、土壤酶及土壤化学性质的影响

花椒（Zanthoxylum bungeanum Maxim.）是川西地区重要的经济植物，化感作用是花椒连作障碍的原因之一，而花椒凋落物和根系分泌物对土壤质量的影响是花椒化感作用的一个重要方面。系统研究花椒如何影响土壤有助于深入理解和解决花椒连作障碍。本文主要以大红袍（10a生）花椒叶和种植过花椒的土壤的浸提液浇灌花椒幼苗进行试验，分析叶浸提液与土壤浸提液对非花椒生长土壤中土壤微生物、土壤酶及土壤化学性质的影响。主要结果如下： 1.花椒叶浸提液和土壤浸提液减少了土壤中微生物的种类、组成和数量。本试验中未施加浸提液的土样中根际微生物明显高于非根际区，在经过花椒叶浸提液处理后，根际细菌、真菌和放线菌数量以及微生物总数都有所减少，这样将会导致土壤中的有效养分的供给减少，进而可能影响植物的生长。 2．施加花椒叶浸提液和土壤浸提液，以及花椒幼苗的栽种，对不同土样中的土壤酶各有促进和抑制作用。在浸提液处理下，水解酶之间及氧化还原酶之间各存在相互促进作用。 3.施加花椒叶浸提液和土壤浸提液均抑制了根际土中全氮和有机质含量，叶浸提液还抑制了无苗土中全磷含量，土壤浸提液还抑制了无苗土中全氮含量与根际土全磷、有机质含量。但两种浸提液均促进了根际土中有效磷和水解性氮含量、根外土中全磷含量，叶浸提液促进了根际土中全磷含量，土壤浸提液促进了根外土中有效磷含量。全氮和有机质含量的下降可能对植物生长发育不利。 4．土壤化学性质与土壤酶活性在不同土样中有不同的相关性。全氮含量在施加叶浸提液的土样中与蛋白酶活性呈正相关。水解性氮含量在施加叶浸提液的土样中与蛋白酶活性、蔗糖酶活性呈正相关。全磷含量在施加叶浸提液的土样中与多酚氧化酶活性呈正相关；在施加土壤浸提液的土样中与蛋白酶活性、蔗糖酶活性呈正相关，与多酚氧化酶活性呈负相关。有效磷含量在施加叶浸提液的土样中与多酚氧化酶活性呈正相关，与蛋白酶活性呈负相关；在施加土壤浸提液的土样中与蛋白酶活性、过氧化氢酶活性呈正相关。有机质含量在施加叶浸提液的土样中与蛋白酶活性、蔗糖酶活性呈正相关。 Zanthoxylum bungeanum is one of the most important cash crops in Eastern Tibetan Plateau. Allelopathic effects could be one of reasons for Z. bungeanum’s continuous cropping impediment. The effects of secretion of leaf and root of Z. bungeanum on soil quality is a important way of Z. bungeanum’s allelopathic effects. However, allelopathic effect of Z.bungeanum on soil microbes, enzyme activities and chemical property were seldom studied. In this study, leaf and soil extracts of Da Hongpao(DHP), the most common varieties of Z.bungeanum in this area, were used to assess allelopathic effect of Z. bungeanum on soil biology and biochemistry by pot experiments . The main results showed that: 1. The irrigation of two kinds of extracts reduced the species, component and quantity of soil microbes. In rhizosphere soil which irrigated by distilled water, the quantity of soil microbes is significantly different from exoroot soil. In rhizosphere soil which irrigated by leaf extracts, the quantity of bacterial, fungi, actionmycete and gross of microbes were decreased, it may resulted in reduce of Available nutrient in soil, and influenced the growth of plants. 2．The irrigation of two kind of extracts reduced or enhanced the enzyme activities in different soils. Interaction between hydrolytic ferments and redoxases were promoted each other. 3. The irrigation of two kinds of extracts reduced the total N and organic matter in rhizosphere soil. Leaf extracts also reduced the total P in soil without seedling. Soil extracts reduced total N in soil without seedling and total P, organic matter in rhizosphere soil. But both extracts also enhanced available P and hydrolysable N in rhizosphere soil, total P in exoroot soil. Leaf extracts enhanced total P in rhizosphere soil. Soil extracts enhanced available P in exoroot soil. The reduction of total N and organic matter may influence growth of plants. 4．Positive correlations between total N and prolease, hydrolysable N and prolease, hydrolysable N and saccharase, total P and polyphenol oxidase, available P and polyphenoloxidase, organic matter and prolease, organic matter and saccharase, were studied in soil irrigated by leaf extracts. In soil irrigated by soil extracts, there are positive correlations between total P and prolease, total P and saccharase, available P and prolease, available P and catalase, while negative correlation between total P and polyphenoloxidase, available P and prolease, available P and catalase was found.

荞麦(Fagopyrum tataricum Gaertn.和F. esculentum Moench.)对紫外线B辐射的响应研究

近二十多年来，基于对臭氧层衰减、紫外线B（UV-B）增强的担心，研究者希望了解到紫外线辐射对不同作物的影响情况，增强UV-B辐射条件下是否对作物的生长发育、产量质量构成威胁。在本试验中，我们首先探讨了双子叶作物黄瓜（Cucumis sativus）和大豆（Glycine max）对不同紫外波段的生物效应[分别为B-UVA（315-400 nm），N-UVA（315-340 nm），B-UVB（275-400 nm）和N-UVB（290-340 nm），UV-（>400nm）作对照]。我们观察到所有的UV波段处理都使黄瓜和大豆的生长受到抑制，并且细胞受到不同程度的氧化伤害；UV波段处理的作用效果与不同波段的紫外有效生物辐射剂量有关。处理差异在UV-B波段内部和UV-A波段内部同样存在。植物生长UV辐射公式（BSWF）能很好的预测本试验UV-B波段内的平均植物效应，但不能预测UV-A波段的植物效应。短波UV-A的生物作用强于长波UV-A。光合色素的变化与UV波谱差异和种间差异有关。在高的紫外/可见光背景下，UV-A处理同UV-B同样导致光合色素的降低，但黄瓜类胡萝卜素/叶绿素比例升高。与其他研究者的试验结果比较后，我们认为紫外线B辐射的生物效应一致性很高，但紫外线A波段的生物学效应存在较大争议。因此我们在本试验的基础上仅进行荞麦[苦荞(Fagopyrum tataricum Gaertn.)和甜荞(Fagopyrum esculentum Moench.)]对紫外线B波段的响应研究。 我们对苦荞品种-圆籽荞进行了连续两个生长季节的大田半控制试验以观察UV-B辐射对苦荞生长、发育、产量及叶片色素的影响；试验小区进行降低UV-B、近充足UV-B和增强UV-B辐射处理。我们的试验表明在不同强度UV-B辐射下苦荞的生长、地上部生物量积累及最终产量都有所下降，但苦荞的发育加快；当前条件下的日光紫外线B辐射对植物生长和产量也造成负面影响。植物光合色素被日光及增强UV-B辐射降低；UV化合物及卢丁含量在中低剂量的UV-B辐射强度下显著升高，但在高剂量的增强UV-B辐射下短期升高后迅速下降。我们的试验表明苦荞是一个对UV-B高度敏感的作物。苦荞对UV-B的敏感性与UV-B剂量、外界环境因素及生长季节有关。 单个苦荞品种的试验结果使我们认识到外界UV-B辐射已经对苦荞生长发育构成逆境条件，未来全球气候变化条件下增强紫外线B辐射可能使其处于更不利的生长环境中。因此我们有进行了多个种群进行UV-B响应观察并筛选耐性种群。我们对15个苦荞种群进行增强UV-B辐射处理（6.30 kJ m2 UV-BBE，模拟当地25%的臭氧衰减），我们观察苦荞UV-B辐射效应存在显著的种内差异，UV-B辐射对多数种群具有抑制作用，但对一些种群还有刺激作用。我们采用主成分分析方法与作物UV-B响应指数（RI）来评价苦荞作物UV-B辐射耐性。我们发现作物的UV-B耐性不仅与其原产地背景UV-B强度有关，而且与作物相对生长效率、次生代谢产物含量（如卢丁）及其他因素有关。我们观察到苦荞伸展叶总叶绿素变化与UV-B耐性成正相关；室内苦荞幼苗的UV-B辐射致死试验表明：苦荞种群死亡率与其UV-B耐性成负相关。 此外，我们对甜荞的UV-B辐射响应也进行了初步研究。选取美姑甜荞、巧家甜荞和云龙甜荞进行5个梯度的增强UV-B辐射室外模拟试验。我们观察到UV-B辐射显著降低了甜荞的生长、生物量及产量；并严重影响了甜荞的生殖生长，降低了花序数、种子数和结实率；并且UV-B辐射对甜荞的抑制作用存在显著的剂量效应。三种甜荞品种存在显著的种内差异，其中美姑品种UV-B耐性最强，且膜脂受UV-B辐射氧化伤害最小，这与该品种UV-B辐射下较高的GR酶活性、APX酶活性和PPO酶活性、以及含量更高的抗坏血酸有关。甜荞的次生代谢也受到增强UV-B辐射的影响，其香豆酰类化合物在UV-B辐射下升高显著，而槲皮素含量也在高剂量UV-B辐射下有所增加；卢丁含量依赖UV-B辐射剂量而变化，中低剂量UV-B辐射下其卢丁含量逐渐升高，但在高剂量辐射下逐渐下降。 通过对生长在高海拔地区的荞麦作物(苦荞和甜荞)进行的室外研究，我们认识到作物不同品种存在很大的耐性差异，这就为UV-B耐性育种创造了有利条件。进一步加大荞麦种质资源筛选力度并深入认识荞麦抗性机理，在此基础上通过杂交或其他基因融合手段培育抗性品种，对高剂量UV-B辐射地区的荞麦产量的提高将起到重要推动作用，并使荞麦生产能有效应对未来全球气候变化条件下UV-B辐射可能升高的威胁。 During last few decades, due to concern of ozone layer depletion and enhancement of ultraviolet B radiation（UV-B, 280-315 nm), the agronomist want to know the responses of different crop species to UV-B. In the first experiment of our study, the effect of different UV band [B-UVA（315-400 nm), N-UVA（315-340 nm), B-UVB（275-400 nm), N-UVB（290-340 nm）and UV-（>400nm, as control)] on the cucumber（Cucumis sativus）and soybean（Glycine max）were investigated in growth room. Spectra-dependent differences in growth and oxidation indices existed within UV-A bands as well as UV-B bands. The general biological effects of different band were UV- ＜ B-UVA＜ N-UVA＜N-UVB＜B-UVB. The plant growth biologically spectra weighting function（BSWF）matched well with average plant response in UV-B region, but not in UV-A region. Shorter UV-A wavelength imposed more negative impact than longer UV-A wavelength did in both species. The effect on photosynthetic pigment was related to different UV bands and different species. The photosynthetic pigment content was decreased by UV-A spectra as well as UV-B spectra. In comparison with the results of previous studies, we found that the wavelength-dependent biological effect of ultraviolet B radiation has high consistency, but the biological effect of ultraviolet-A radiation was inconsistent. We narrow our following study on the effect of ultraviolet B radiation on the buckwheat（tartary buckwheat and common buckwheat). The tartary buckwheat（Fagopyrum tataricum Gaertn.）cultivars Yuanziqiao was grown in the sheltered field plots for two consecutive seasons under reduced, near-ambient and two supplemental levels of UV-B radiation. The crop growth, photosynthetic pigments, total biomass, final seed yield and thousand-grain weight were decreased by near-ambient and enhanced UV-B radiation, while crop development was promoted by enhanced UV-B radiation. Leaf rutin concentration and UV-B absorbing compound was generally increased by UV-B with the exception of 8.50 kJ m-2 day-1 supplemental levels. Our results showed that tartary buckwheat is a potentially UV-B sensitive species. Study on one cultivars showed that ambient solar radiation had present a stress to tartary buckwheat. This makes it necessary to observe the UV-B response of many cultivars and screen tolerant cultivars. Fifteen populations of tartary buckwheat were experienced enhanced UV-B radiation simulating 25% depletion of the stratospheric ozone layer in Kunming region, and plant responses in growth, morphology and productivity were observed. Principal components analysis（PCA）was used to evaluate overall sensitivity of plant response to UV-B as well as response index. The different populations exhibited significant differences in responses to UV-B. The photosynthetic pigments of young seedlings were also affected significantly under field condition. On the other hand, the healthy seedlings of different populations were exposed to the high level of UV-B radiation in growth chambers to determine the plant lethality rate. The plant tolerance evaluated by multivariate analysis was positively related to total plant chlorophyll change, but negatively related to lethality rate. In other hand, the UV-B responses of the other important cultivated buckwheat species, common buckwheat（Fagopyrum esculentum Moench.), were also studied preliminarily. Three widespread cultivated variety（Meigu, Qiaojia and Yunlong cultivars）were provided with five level of enhanced UV-B radiation outdoors. We observed that the crop growth, development and production were significantly decreased, and reproductive production, like anthotaxy number, seed number and seed setting ratio, was also decreased. Dose-dependent inhibition effect caused by enhanced UV-B radiation also existed in common buckwheat. Significant intraspecific difference existed in those three cultivars. The Meigu cultivars with dwarfed growth and lower production have highest UV-B tolerance as well as lowest damage in cell membrane, this could be associated with profound enhancements of glutathione reductase（GR）activity, ascorbate peroxidase activity and polyphenol oxidase activity as well as higher ascorbic acid concentration. The secondary metabolism was also affected by UV-B radiation, with profound elevation of coumarin compound and moderate increase of quercetin concentration. Rutin concentration was peaked in 5kJ m-2 UV-B. The contrasting effect of UV-B radiation on different populations indicated that there existed abundant genetic resources for selecting tolerant populations of common and tartary buckwheat. Much effort needed be pose on screening of buckwheat germplasm and clarification of mechanism of buckwheat tolerance to UV-B. On this base the tolerant cultivars could be bred by hybridization and other gene transfusion method, this would help increase buckwheat yield in high ambient UV-B region and counteract the effect of possible enhanced UV-B radiation in future.

青杨组(Section Tacamahaca Spach)杨树不同种群对土壤干旱和重金属锰污染的生态生理响应

土壤是人类赖以生存的自然环境和农业生产的重要资源，世界面临的粮食、资源和环境问题与土壤密切相关，目前危害土壤的主要因素是干旱和重金属污染。杨树具有适应性强、生长快和丰产等特性，本论文以青杨组杨树为模式植物，采用植物生态、生理及生物化学等方法，研究杨树对土壤干旱和锰胁迫的生态生理反应以及种群间差异，研究成果可为我国干旱半干旱地区营造人工林、防止沙漠化提供理论依据，也为恢复与重建重金属污染地区退化生态系统提供科学指导。主要研究结果如下： 1. 青海杨不同种群对干旱胁迫的响应差异 干旱胁迫显著降低了两个青海杨种群（干旱种群和湿润种群）生物量积累，包括株高、基径、干物质积累等，通过植物结构的调整，有更多的生物量向根部分配。干旱胁迫还显著降低了叶绿素和类胡萝卜素含量，增加了游离脯氨酸和总氨基酸含量。另一方面，干旱胁迫诱导了活性氧的积累，作为第二信使，激活了抗氧化系统，包括抗坏血酸（ASA）含量和酶系统如超氧化物歧化酶（SOD），愈创木酚过氧化物酶（GPX），抗坏血酸过氧化物酶（APX）和谷胱甘肽还原酶（GR）。这样，杨树既有避旱机制又有耐旱机制，使其在干旱胁迫下有相当程度的可塑性。与湿润种群相比，干旱种群杨树有更多的生物量分配到根部，积累了更多的游离脯氨酸和总氨基酸来进行渗透调节，并且有更有效的抗氧化系统，包括更高含量的ASA 和更高活性的APX 和GR，这些使得干旱种群杨树比湿润种群杨树对干旱有更好的耐性。 2. 喷施硝普钠（SNP）对青海杨阿坝种群干旱胁迫耐性的影响 干旱胁迫显著的降低了青海杨阿坝种群的生长和生物量积累以及叶片相对含水量，还诱导了脯氨酸的合成以进行渗透调节。干旱胁迫下过氧化氢（H2O2）显著累积从而造成对膜脂和蛋白的伤害，使得丙二醛和蛋白羰基含量升高。干旱胁迫下喷施SNP可以减轻干旱胁迫造成的伤害，包括增加叶片的相对含水量，增加脯氨酸和总氨基酸的积累，并激活抗氧化酶系统如SOD，GPX和APX，从而减少丙二醛（MDA）和蛋白羰基（C＝O）的积累，但是在水分良好情况下SNP的效果不显著。 3. 青杨不同种群对锰胁迫的生长与形态响应差异 在同一锰浓度下，干旱种群的耐性指数都要高于湿润种群，这表明青杨对干旱和高锰胁迫具有交叉耐性。两个种群的株高，生物量和叶绿素含量都随锰浓度的升高而逐渐下降。就累积浓度而言，0 和0.1 mM 锰胁迫下，干旱种群积累的锰浓度要高于湿润种群，而在高浓度锰胁迫下(0.5 和1 mM)，湿润种群要高于干旱种群。在0，0.1 和0.5 mM下，锰大多积累在根中，叶片次之，茎中最少。而在1 mM，锰更多的积累在叶片中。就累积总量而言，在各个锰浓度胁迫下，根，茎和叶相比，两个种群青杨都是叶片累积的锰总量要高于根和茎。两个种群间比较，对照中没有显著区别，0.1 mM 锰胁迫下，湿润种群根中累积的锰要高于干旱种群，而在地上部中，干旱种群要高于湿润种群。而0.5 和1 mM 锰胁迫下，根、叶、茎+叶、根+茎+叶中，锰累积总量都是湿润种群高于干旱种群。锰胁迫下，青杨叶片数和叶面积包括总叶面积和平均叶面积都显著降低。叶片横切面的光学显微观察结果表明未经锰胁迫的栅栏组织的细胞饱满，海绵组织发达、清晰；胁迫后杨树叶片栅栏组织细胞出现不同程度的皱缩，海绵组织几乎不可见，此外还发现输导组织在胁迫下密度变小和分生组织严重割裂等现象。 4. 青杨不同种群对锰胁迫的生理与生化响应差异 青杨两个种群脱落酸（ABA）含量在锰胁迫下都显著增加，干旱种群的增幅更大。三种多胺含量在锰胁迫下显示了不同的响应趋势：腐胺在两个种群的各个锰处理下都增加，亚精胺只在干旱种群中显著增加，而精胺除了干旱种群在1 mM 下有所增加外，在锰胁迫下变化很小。谷胱甘肽含量随锰浓度升高而增加，在0.5 mM 锰时达到最高值，1mM 时有所下降。植物络合素（PCs）含量与非蛋白巯基（NP-SH）趋势相似，随锰浓度的升高而增加，且干旱种群中含量要高于湿润种群。锰处理还引起氧化胁迫，表现为过氧化氢和丙二醛含量增加。SOD 活性在湿润种群中，在0 到0.5 mM 锰胁迫下活性升高，但在1 mM 锰胁迫时，其活性有所下降。而在干旱种群中，SOD 活性变化较小，并始终维持在一个较高的水平。APX 活性在两个种群中都随锰浓度的升高而增加，干旱种群活性要高于湿润种群。锰胁迫还显著增加了酚类物质的含量，同时GPX 和多酚氧化酶（PPO）活性也随锰浓度的升高而增加。干旱种群的酚类含量和GPX 与PPO 活性都要高于湿润种群。锰胁迫还改变了氨基酸的含量和构成，根据锰胁迫下浓度变化的不同，可以将游离氨基酸分为三组：第一组包括，谷氨酸，丙氨酸和天门冬氨酸，这一组氨基酸含量在锰胁迫下有所下降。第二组包括缬氨酸，亮氨酸和苏氨酸含量在锰胁迫下基本不变化或变化很小。剩下的氨基酸为第三组，这组氨基酸含量在锰胁迫下显著增加，而根据增加的幅度又可以将它们分为两个亚组，丝氨酸，酪氨酸，苯丙氨酸，组氨酸和脯氨酸，在1 mM 下的含量是对照的4 倍以上。异亮氨酸，赖氨酸，精氨酸和甘氨酸含量在1 mM 下是对照含量的2 倍以下。同时，同一锰浓度下，干旱种群比湿润种群积累的氨基酸含量要高。 Soil is the indispensable environment for human survival and important resource foragriculture development. Food and environmental problems facing the world are all closelyrelated to soil and nowadays it is threatened by many factors, among which drought stress andheavy metal pollution are the most serious ones. Poplars (Populus spp.) are importantcomponents of ecosystem and suitable as a source of fuel, fiber and lumber due to their fastgrowth. In this study, different populations of Section Tacamahaca spach were used as modelplants to investigate the adaptability to drought stress and manganese toxicity and differencesbetween populations from dry and wet climate regions. Our results can provide theoreticalevidence for the afforestation and prevention of desertification in the arid and semi-arid areas,and also can supply scientific direction for the reconstruction and rehalibitation of ecosystemscontaminated by heavy metals. The results are as follows: 1. Differences in ecophysiological responses to drought stress in two contrastingpopulations of Populus przewalskii Drought stress not only significantly affected dry mass accumulation and allocation, butalso significantly decreased chlorophyll pigment contents and accumulated free proline andtotal amino acids. On the other hand, drought also significantly increased the levels ofabscisic acid and reactive oxygen species, as secondary messengers, to induce the entire set ofantioxidative systems including the increase of reduced ascorbic acid content and the activities of superoxide dismutase, guaiacol peroxidase, ascorbate peroxidase and glutathioneredutase. Thus the combination of drought avoidance and tolerance mechanisms conferredpoplar a high degree of plasticity in response to drought stress. Compared with the wetclimate population, the dry climate population showed lower dry matter accumulation andallocated more biomass to root systems, and accumulated more free proline and total aminoacids for osmotic adjustment. The dry climate population also showed more efficientantioxidant systems with higher content of ascorbic acid and higher activities of ascorbateperoxidase and glutathione redutase than the wet climate population. All these made the dryclimate population superior in adaptation to drought stress than the wet climate population. 2. Effect of exogenous applied SNP on drought tolerance in Populus przewalskii Drought stress significantly increased hydrogen peroxide content and caused oxidativestress to lipids and proteins assessed by the increase in malondialdehyde and total carbonylcontents, respectively. The cuttings of P. przewalskii accumulated proline and other aminoacids for osmotic adjustment to lower water potential, and activated the antioxidant enzymes such as superoxide dismutase, guaiacol peroxidase and ascorbate peroxidase to maintain thebalance of generation and quenching of reactive oxygen species. Moreover, exogenous SNPapplication significantly heightened the growth performance of P. przewalskii cuttings underdrought treatment by promotion of proline accumulation and activation of antioxidant enzymeactivities, while under well-watered treatment the effect of SNP application was very little. 3. Morphological responses to manganese toxicity in the two contrasting populations ofPopulus cathayana High concentration of manganese caused significant decrease in shoot height andbiomass accumulation. The tolerance index of the dry climate population was significantlyhigher than that of the wet climate population, suggesting the superior Mn tolerance in theformer and the existence of cross-tolerance of drought stress and high Mn toxicity. Injuries tothe leaf anatomical features were also found as the reduced thickness in palisade and spongyparenchyma, the decreased density in the conducting tissue and the collapse and split in themeristematic tissue in the central vein. As for the Mn concentrations in the plant tissues, under0, 0.1 and 0.5 mM, most of the Mn accumulated in the roots, then leaves, and stem the least, while under 1 mM, most of the Mn accumulated in the leaves. As far as the total amounts ofMn extraction are concerned, the leaf extracted more Mn than the root and stem in the twopopulations under various Mn concentrations. There is no difference between the twopopulations under control. Under 0.1 mM, the wet climate population extracted higher Mn inthe root than the dry climate population, while in the shoot, the dry climate populationextracted much more Mn. Under 0.5 and 1 mM, the wet climate population translocated moreMn both in the root and the shoot than the dry climate population. 4. Physiological and biochemical responses to manganese toxicity in the two contrastingpopulations of Populus cathayana Mn treatment resulted in oxidative stress indicated by the oxidation to lipids, proteinsand DNA. A regulated network of defence strategies was employed for the chelation,detoxification and tolerance of Mn including the enhanced synthesis of ABA and polyamines,the accumulation of free amino acids, especially His and Pro, and the activation of theenzymes superoxide dismutase and guaiacol peroxidase. Contents of non-protein thiol,reduced glutathione, phytochelatins and phenolics compounds and activities of superoxide dismutase, guaiacol peroxidase and polyphenol oxidase also increased significantly forantioxidant or chelation functions. The wet climate population not only accumulated lessabscisic acid, free amino acids, phytochelatins and phenolics compounds, but also exhibitedlower activities of superoxide dismutase, guaiacol peroxidase and polyphenol oxidase thusresulting in more serious oxidative damage and more curtained growth.

模拟增温对川西亚高山人工云杉林土壤碳含量和土壤呼吸的影响

全球气候变化已经成为不争的事实，其中全球变暖是近年来国内外的研究热点之一。土壤碳库作为陆地生态系统最大的碳库，气温升高必然会导致一系列的土壤碳储量和碳通量的变化，这些微小的变化又可能导致大气CO2浓度的变化并强化这种变暖的趋势。目前，土壤碳循环对温度升高的响应仍然是陆地碳循环研究最缺乏的部分，对土壤有机碳动态变化的研究仍存在着很大的不确定性与争议。四川西部的亚高山人工针叶林是青藏高原东部高寒林区的重要组成部分，是研究全球变化对森林生态系统影响的关键地区和重要森林类型。本研究通过采用原位人工模拟增温装置（Open-top chambers，OTCs）对川西米亚罗60年人工云杉林土壤实施增温，研究高海拔地区森林，尤其是人工森林系统下的土壤有机碳 含量、土壤呼吸及土壤酶活性对温度升高的响应。结果表明： 1. 增温处理的660天（2005年11月至2007年9月）期间，增温条件下的平均气温和土壤平均温度分别比对照提高0.43 ℃和0.27 ℃；0~10 cm土壤含水量在增温的不同时期均有不同程度的降低。 2. 土壤蔗糖酶、蛋白酶和脲酶活性在温度升高的不同阶段均有不同程度的提高。在增温处理300天（2006.09）、540天（2007.05）、600天（2007.07）和660天（2007.05）后，0~10 cm层的蔗糖酶活性分别比对照提高了36.36%（P<0.05）、24.31%、14.54%（P<0.05）和7.22%，脲酶活性分别提高了12.90%、24.19%（P<0.01）、34.48%（P<0.05）和14.64%(P<0.05)，蛋白酶活性分别提高了31.37%、1.99%、3.70%和17.80%。10~20 cm层的土壤酶活性也均有不同程度的提高，但均没有显著差异。蔗糖酶、脲酶和蛋白酶活性均呈现出随土层加深而减弱的趋势。 3. 土壤过氧化氢酶和多酚氧化酶活性在增温的第1年内均有不同程度的提高，但在增温的第2年内比对照有所降低。增温300天后（2006.09），过氧化氢酶和多酚氧化酶在0~10 cm层分别比对照增加3.76%和49.25%（P<0.05），10~20 cm层分别增加了5.54%和29.67%。在增温的第2年内，增温540天（2007.05）、600天（2007.07）和660天（2007.09）后，0~10 cm层的过氧化氢酶活性分别比对照降低了27.70%（P<0.05）、4.34%和1.47%，多酚氧化酶活性分别降低了5.86%、11.76%（P<0.05）和7.47%。增温的第2年内，10~20 cm层的过氧化氢酶和多酚氧化酶活性也均有不同程度的降低，但差异均未达到显著水平。不同土层之间相比较，过氧化氢酶活性随土层加深而降低，多酚氧化酶活性随土层加深而增加。 4. 土壤有机碳和有机质在增温的不同阶段，含量比对照均有所降低；且随增温时间的延长，降低的幅度下降。0~10 cm层的土壤有机碳和土壤有机质在增温300天（2006.09）、540天（2007.05）、600天（2007.07）和660天（2007.09）后分别降低了8.69%、4.35%、3.80%和2.44%，差异均未达到显著水平。土壤全氮含量在增温后与对照相比无明显的增加或者降低趋势。增温条件下的土壤C/N比与对照相比有所降低，但在增温各阶段的差异均不显著。10~20 cm层的有机碳、有机质和C/N比也有不同程度的降低趋势，但差异均不显著。不同土层之间相比，0~10 cm层的有机碳、有机质、全氮含量和C/N比均高于10~20 cm层，呈现出随土层加深而降低的趋势。 5. 土壤呼吸速率在增温第1年内，与对照相比明显提高，但在增温处理2年后，与对照相比无显著变化。增温300天（2006.09）和360天（2006.11）后分别提高了13.32%和21.17%，差异显著。增温处理540天（2007.05）到660天（2007.09）期间，与对照相比，不仅没有明显的提升，反而有些月份比对照有所降低，对温度升高的敏感性降低，呈现出对温度升高的适应性。土壤呼吸的日呼吸速率呈现单峰曲线形式，在14:00~20:00期间达到最大值，在4:00~10:00期间具有最低值。土壤呼吸的季节变化，呈现出与外界环境温度相一致的趋势，在7月份（夏季） 最高，11月份（冬季）最低。土壤呼吸与2 cm土壤温度、5 cm土壤温度和空气温度均呈极显著指数相关，与0~10 cm土壤含水量呈线性相关，相关性达到显著水平，但低于土壤呼吸与温度的相关性。 The past century has seen a marked increase in atmospheric carbon dioxide concentrations and a concomitant warming that has drawn scientific attention to the link between global carbon stocks and climate change. In particular, the decomposition and turnover of soil organic matter is recognised as an important determinant of carbon driven climate change. The slightly variation in soil organic carbon will result in the increase of atmospheric carbon dioxide concentrations and reinforce the tendency of warming. The experiment was conducted in Subalpine coniferous forest in western Sichuan province. Subalpine coniferous forest in western Sichuan was a important part of eastern Qinghai-Tibetan Plateau, which play a important role in reseaching the sensitivity of forest ecosystem to climate change. To investigate the effects of elevated temperature on soil organic carbon content, soil respiration rates, and soil enzyme activities in subalpine Picea asperata plantations, a esimulated warming measure was applied with Open-top chambers. The results were as followed: 1) During the period from Nov. 2005 to Sep. 2007, mean air temperature and soil temperature were respectively 0.43℃ and 0.27℃ the ambient higher. Soil water content decreased to different exent in different months in warmed plots than in unwarned plots at depth of 0-10 cm. 2) In general, elevated temperature enhanced the soil enzyme activities of invertase, protease, and urease. In the first year of warming—after 300 days’ treatment (in Sep,2006), the activities of invertase, protease, and urease increased by 36.36%, 12.90% and 31.37% respectively at the depths of 0-10 cm，among which the activity of invertase reached statistic significance. In the second year of warming, invertase activity increased by 24.31% after 540 days’ treament (in May, 2007), 14.54% after 600 days’ treament (in Jul, 2007) and 7.22% after 660 days’ treatment (in Sep, 2007) at the depths of 0-10 cm, and the differences in July and Septemmber were statistically significant. Elveated temperature also increased the activity of urease in the second year of warming and had significant effects in May and July. The activity of protease in warmed plots was also higher than in unwarmed plots at depths of 0-10 cm, but there was no significant difference. Elevated temperature had no significant effects on all soil enzyme acitivities at the depths of 10-20 cm in the first and sencond year. The values of above-mentioned soil enzyme all decreased with soil layers. 3) Eleavted temperature enhanced the activities of catalase and polyphenol oxidase in the first year of warming while they turned out downtrend in the second year. The activity of catalase increased by 3.76% and 5.54% at depths of 0-10 cm and 10-20 cm respectively in the first year—after 300 days’ warming (in Sep, 2006), the differences of which had no statistical significance. The activity of polyphenol oxidase was significantly increased by 49.25% at depths of 0-10 cm and not significantly increased by 29.67% at depths of 10-20 cm after 300 days’ warming. In the second year of warming, the catalase activity was significantly decreased by 27.70% after 540 days’ treament (in May, 2007) and not significantly decreased by 4.34% and 1.47% after 600 days’ (in Jul, 2007) and 660 days’ treament (in Sep, 2007) respectively. The activities of catalase and polyphenol oxidase at depths of 10-20 cm were decreased to different extent, but there was no significant difference. Catalase activity stepped down with soil layers while polyphenol oxidase activity stepped up. 4) Increased temperature in both the first year and the second year resulted tendency of decrease in the contents of soil organic carbon and soil organic matter, and C/N ratios at soil depths of 0-10 cm and 10-20 cm. However, with the prolonged warming, the tendency of decrease gradually tapered off and the extent of decrease in the second year of experiment were lower than that in the first year. The contents of soil organic carbon and soil organic matter were all decreased 8.69% by warming in the first year and dcreased 4.35%, 3.80% and 2.44% in May, July and September of the second year, but no significant difference were found. The C/N ratios increased 8.52% in the first year of warming and had less increment in the second year, all of which were not statistical significant. Eleveated temperature had no obvious effect on the content of tatol N in two year consecutive warming experiment. The contents of soil organic carbon and soil organic matter, total N and C/N ratios all had the tendency of dcreasing with soil layers. 5) Soil respiration rates were significantly enhanced by 13.32% and 21.17% after 300 days’ (in Sep, 2006) and 360 days’ (in Nov, 2006) treament in the first year of warming, but the same showed no obvious difference in the second year of treatment, which was assumed the adaptability of soil respiration with a certain heightened temperature. Diurnal soil resspiration showed a daily variation with a minimum value between 4:00 and 10:00 h and a maximum value between 14:00 and 20:00 h, coinciding with the minimum and maximum values of soil temperature at 2 cm. Soil respiration rates exhibited a pronounced seasonal variation with minimum values in Novmber and a maximum value in July, approximately coinciding with the seasonal variation of air and soil temperature. An exponential function provided the best fit for soil respiration with temperature while a quadric equation was used to estimate the effect of soil moisture on soil respiration, which were all significantly correlated. Soil respiraion rate was more highly correlated with the soil temperature than soil moisture.

鲜甘薯快速乙醇发酵条件的初步研究

本文对不同菌种（酵母菌和运动发酵单胞菌）快速生产燃料乙醇的条件进行了研究，实现了鲜甘薯快速转化为燃料乙醇。全文分为两部分： 第一部分：酵母菌快速生产燃料乙醇的条件研究。通过单因素试验，酵母菌快速生产燃料乙醇的条件为：发酵方式采用边糖化边发酵（SSF），蒸煮温度为85 ℃，料水比2:1（初始糖浓度 210 g/kg），糖化酶用量0.75 AGU/g 鲜甘薯，接种量10%（v/w）。在最优条件下，经过24 h发酵，乙醇浓度可达97.44 g/kg, 发酵效率为92%，发酵强度为4.06 g/kg/h。由于采用了低温蒸煮和SSF，可以大大节约能耗，从而降低乙醇生产的成本。同时，利用摇瓶优化的条件，进行了10 L，100 L，500 L发酵罐的放大试验，由于发酵罐初期可以人为通氧，使菌体能迅速积累，发酵时间缩短2 h，发酵效率在90%以上。 第二部分：运动发酵单胞菌快速生产燃料乙醇条件研究。通过单因素试验和正交试验获得了发酵的最佳参数：初始pH值6.0-7.0，硫酸铵5.0 g/kg，糖化酶量1.6 AUG/kg淀粉，初始糖浓度200 g/kg，接种量12.5%（v/w）。经过21 h发酵，乙醇浓度为95.15 g/kg，发酵效率可达94%。同时对不灭菌发酵也进行了研究，发酵效率可达92%。为鲜甘薯运动发酵单胞菌燃料乙醇的工业化生产打下基础。 对发酵结束后的残糖进行了研究。通过薄层层析和葡萄氧化酶测定证明：无论是酵母菌还是运动发酵单胞菌发酵结束后的发酵液中都不含葡萄糖。经过HPLC进一步分析残糖说明：发酵液中已没有葡萄糖成分；经糖化酶水解后仍没有葡萄糖出现；但经酸水解后又出现了葡萄糖，说明结束后的残糖是一些低聚糖结构。有关残糖的结构需要进一步研究。可以通过开发高效的低聚糖水解酶来降低发酵液的残糖，提高原料的利用率。 A new technology for rapid production fuel ethanol from fresh sweet potato by different microorganisms (Saccharomyces cerevisiae and Zymomonas mobilis) was gained in this research. The paper involved two parts: Part 1: The study on fuel ethanol rapid production from fresh sweet potato by Saccharomyces cerevisiae. The following parameters of Saccharomyces cerevisiae was investigated by a series of experiments: fermentation models, cooking temperature, initial sugar concentration and glucoamylase dosage. The results showed that SSF (simultaneous saccharification and fermentation) not only reduced the fermentation time (from 30 to 24h) but also enhanced the ethanol concentration (from 73.56 to 95.96 g/kg). With low-temperature-cooking (85 ℃) using SSF, the Saccharomyces cerevisiae was able to produce ethanol 97.44 g/kg which the fermentation yield could reach to 92% and ethanol productivity 4.06 g/kg/h from sweet potato enzymatic hydrolysis. Furthermore, the savings in energy by carrying out the cooking (85 ℃) and saccharification (30 ℃) step at low temperature had been realized. The results were also verified in 10 L, 100 L and 500 L fermentor. The fermentation yield was no less than 90%. The fermentation time of fermenter was shorter than Erlenmeyer flask. This may be that the aeration in the early fermentation period is available, which lead to the rapidly commutations of biomass. Part 2: The technology of ethanol rapid production with simultaneous saccharification and fermentation ( SSF ) by Zymomonas mobilis，using fresh sweet potato as raw material was studied. The effects of various factors on the yield of ethanol were investigated by the single factor and the orthogonal experiments. As a result, the optimal technical conditions were obtained from those experiments：initial pH value 6.0-7.0, nitride 5.0 g/kg，(NH4)2SO4, glucoamylase 1.6 AUG/kg starch, inoculums concentration 12.5% (v/w). The Zymomonas mobilis was able to produce ethanol 95.15 g/kg, with 94% of the theoretical yield, from fresh sweet potato after 24 h fermentation. The fermentation efficiency of non-sterilized was also reach to 92%. We also analyzed the final fermentation residual sugars of Saccharomyces cerevisiae and Zymomonas mobilis. When the residual sugars were analyzed by thin-layer chromatogram and glucose oxidase, there was no glucose. The analysis of reducing sugars by HPLC showed that there was no glucose existed in the fermentation liquor. However, the glucose appeared after being hydrolyzed by acid. It is indicated that the residual sugars in the final fermentation liquor were the configuration of oligosaccharide, which was linked by the special glycosidic bonds. It was feasible for reducing residual sugars to develope the enzyme that can degradation the oligosaccharide.

Organic phase enzyme electrodes based on organohydrogel

A dimethylformamide-polyhydroxyl cellulose organo-hydrogel has been prepared, and its applications for enzyme immobilization in construction of organic phase biosensors have been exploited. With horseradish peroxidase, tyrosinase, and bilirubin oxidase immobilized in the organohydrogel, enzyme electrodes can be operated in various situations, including aqueous buffer, oil/water mixtures, and anhydrous organic solvents, and even in dimethylformamide, to determine analytes of different solubilities, e.g., organic peroxides, phenolic compounds and bilirubin. Biosensing has no restrictions in terms of measuring media and solubilities of analytes.