Water Balance in Cucumis Plants, Measured by Nuclear Magnetic Resonance, II

Nuclear magnetic resonance (NMR) was used to investigate theeffects of changes in root temperature, of changes in the areaof root in contact with culture solution and of day/night rhythmon the water balance of a cucumber and a gherkin plant. Resultsare discussed in terms of water potential, flow rate and resistanceusing a previously presented model of water balance. As longas water uptake alone is varied, flow rate and water content(or potential) will change in the same direction. In contrast,from that model it is predicted that changes in transpirationwill affect flow rate and water content in opposite ways. Anexperimental verification of this prediction was given in theprevious paper. Results obtained by the NMR method are comparedto those determined using a dendrometer. The results demonstratethat the NMR method is a valuable tool to study plant waterbalance and that it can serve as a technique for discriminatingbetween changes in plant water balance that are due to changesin water uptake by roots and those due to changes in transpiration. Key words: Water balance model, Cucumis satious L., flow, water content, NMR, water balance measurement     

Water Balance in Cucumis Plants, Measured by Nuclear Magnetic Resonance, I

In this paper we demonstrate the study of plant water balanceby the non-invasive measurement of tissue water content andwater flow using proton nuclear magnetic resonance (NMR). Sapvelocity and flux were measured independently in the presenceof an excess of stationary tissue water. The instrumentationdescribed allows automated and unattended measurement of flow-and water content-variables in a well-defined region of theplant over periods of several days, with a time resolution betweensuccessive measurements of c. 5 s. Using this apparatus theeffect of changes in light intensity (day/night rhythm) andrelative humidity on stem tissue water content as well as onthe velocity and flux of xylem sap in the stem were investigatedin a cucumber plant. The results are in agreement with predictionsfrom a simple model for plant water balance, which is basedon water potential, flow rate and resistance to flow. As longas only transpiration is varied, flow rate and water content(or potential) are affected in opposite ways as demonstratedin this paper. In contrast, the model predicts that changesin uptake (resulting from changes in, for example, root resistance)will induce changes in water content and flow in the same direction.An experimental verification of this prediction is given ina subsequent paper, where, in addition, the NMR results arecompared to those obtained with a dendrometer. Key words: Water balance model, Cucumis sativus L., flow, water content, NMR, water balance measurement     

Quantification of water transport in plants with NMR imaging

A new nuclear magnetic resonance imaging (NMRi) method is described to calculate the characteristics of water transport in plant stems. Here, dynamic NMRi is used as a non-invasive technique to record the distribution of displacements of protons for each pixel in the NMR image. Using the NMR-signal of the stationary water in a reference tube for calibration, the following characteristics can be calculated per pixel without advance knowledge of the flow-profile in that pixel: the amount of stationary water, the amount of flowing water, the cross-sectional area of flow, the average linear flow velocity of the flowing water, and the volume flow. The accuracy of the method is demonstrated with a stem segment of a chrysanthemum flower by comparing the volume flow, measured with NMR, with the actual volumetric uptake, measured with a balance. NMR measurements corresponded to the balance uptake measurements with a rms error of 0.11 mg s(-1) in a range of 0 to 1.8 mg s(-1). Local changes in flow characteristics of individual voxels of a sample (e.g. intact plant) can be studied as a function of time and of any conceivable changes the sample experiences on a time-scale, longer than the measurement time of a complete set of pixel-propagators (17 min).     

Functional Imaging of Plants: A Nuclear Magnetic Resonance Study of a Cucumber Plant

Functional magnetic resonance imaging was used to study transients of biophysical parameters in a cucumber plant in response to environmental changes. Detailed flow imaging experiments showed the location of xylem and phloem in the stem and the response of the following flow characteristics to the imposed environmental changes: the total amount of water, the amount of stationary and flowing water, the linear velocity of the flowing water, and the volume flow. The total measured volume flow through the plant stem was in good agreement with the independently measured water uptake by the roots. A separate analysis of the flow characteristics for two vascular bundles revealed that changes in volume flow of the xylem sap were accounted for by a change in linear-flow velocities in the xylem vessels. Multiple-spin echo experiments revealed two water fractions for different tissues in the plant stem; the spin-spin relaxation time of the larger fraction of parenchyma tissue in the center of the stem and the vascular tissue was down by 17% in the period after cooling the roots of the plant. This could point to an increased water permeability of the tonoplast membrane of the observed cells in this period of quick recovery from severe water loss.     

不同膜下滴灌方式对设施黄瓜生理特性及水分利用效率的影响

为了探明黄瓜膜下分根交替滴灌的节水效果,为设施黄瓜节水灌溉提供理论依据和技术参数,以‘津优3号’黄瓜为试材,采用随机区组设计,以土壤田间持水量的65%为灌水下限,田间持水量的90%为灌水上限,研究了分根交替滴灌(APDI)、固定1/2根区滴灌(FPDI)和传统滴灌(CDI)3种灌溉模式对黄瓜生长、生理特性、产量与品质及水分利用效率的影响,结果表明:(1)随灌溉处理时间的延长,3种灌溉模式的单株叶面积和株高的差异越来越显著,而茎粗和叶片数差异不显著;(2)与传统滴灌相比,分根交替滴灌模式下黄瓜叶片净光合速率略有下降而蒸腾速率显著降低,水分利用效率显著提高;(3)分根交替滴灌处理下黄瓜可溶性蛋白、可溶性糖含量与传统滴灌相比差异不显著,Vc含量却显著增加;(4)分根交替滴灌模式下黄瓜产量比传统滴灌下降1.5%,而灌水量减少17%,水分利用效率提高18.6%,节水效果显著。综上所述,分根交替滴灌可以在保证设施黄瓜产量没有显著下降的前提下,改善品质和显著提高水分利用效率,可作为设施节水提质增效的一种灌溉模式,推广应用前景广阔。     

Intact plant magnetic resonance imaging to study dynamics in long-distance sap flow and flow-conducting surface area

Due to the fragile pressure gradients present in the xylem and phloem, methods to study sap flow must be minimally invasive. Magnetic resonance imaging (MRI) meets this condition. A dedicated MRI method to study sap flow has been applied to quantify long-distance xylem flow and hydraulics in an intact cucumber (Cucumis sativus) plant. The accuracy of this MRI method to quantify sap flow and effective flow-conducting area is demonstrated by measuring the flow characteristics of the water in a virtual slice through the stem and comparing the results with water uptake data and microscopy. The in-plane image resolution of 120 x 120 microm was high enough to distinguish large individual xylem vessels. Cooling the roots of the plant severely inhibited water uptake by the roots and increased the hydraulic resistance of the plant stem. This increase is at least partially due to the formation of embolisms in the xylem vessels. Refilling the larger vessels seems to be a lengthy process. Refilling started in the night after root cooling and continued while neighboring vessels at a distance of not more than 0.4 mm transported an equal amount of water as before root cooling. Relative differences in volume flow in different vascular bundles suggest differences in xylem tension for different vascular bundles. The amount of data and detail that are presented for this single plant demonstrates new possibilities for using MRI in studying the dynamics of long-distance transport in plants.     

土壤水分对温室春黄瓜苗期生长与生理特性的影响

以津优1号黄瓜为试验材料．研究了土壤不同水分含量对温室春黄瓜苗期生长与生理特性的影响。结果表明,在土壤水分为田间持水量90％时．黄瓜幼苗长势健壮,茎粗大,根系活力强,叶绿素含量和光合速率高．与其它处理差异显著,说明土壤含水量过高或过低均不利于培育黄瓜壮苗。株高和叶面积随土壤含水量的升高而增加;细胞汁液浓度随土壤含水量的升高而降低．且对植株水分状况反应十分敏感,可以用来作为判断植株水分盈亏状况的生理指标。     

An Analysis of the Mussell and Morre Quantitative Bioassay for Polygalacturonases using Pectate trans-eliminase from Erwinia atroseptica

The Mussell and Morré method for estimating the activityof chain-splitting pectic enzymes was tested with sections ofparenchyma from cucumber fruit and potato tubers. It provedto be a very sensitive test for the pectate trans-eliminaseof Erwinia atroseptica, between 10 and 100 times as sensitiveas a viscometric method based on degradation of polypectate.Loss of fresh weight was readily detected within a few minutesafter immersion in enzyme solution and continued until the sectionof tissue had lost coherence. For some time after immersion,loss of weight was mainly due not to loss of cells from thesurface of the section but to loss of water and solutes, presumablybecause of the effect of the trans-eliminase in decreasing thepermeability of the protoplasts. In the later stages loss ofcells became more important. Potato sections lost weight atabout half the rate of cucumber sections but the rate of increaseof absorbance at 235 nm of the ambient solution was about athird higher for potato than for cucumber. Potato sections lostelectrolytes much more rapidly than did cucumber sections. Thesedifferences probably reflect, at least in part, the much higherwater content of cucumber sections. The advantages of the Mussell and Morré method are itssensitivity and the fact that the natural substrates of chain-splittingenzymes can be used for assessing activity.     

Water stress in cucumber cotyledons infected withXanthomonas campestris pv.cucurbitae

Xanthomonas compestris pv.cucurbitae strain 62 was isolated from diseased cucumber seedlings grown in an open field in Egypt and identified in this study. Inoculation of cucumber seedlings with this strain resulted in reduction of relative water content (RWC) and increase in free proline content of infected cotyledons. RWC was found to be decreased by the increase of disease severity. The reverse was true for free proline content which was found to be increased by the increase of disease severity. These results indicate that water stress increased in direct proportion to the disease severity. Highly significant increase in free proline content was found to occur in organs of cucumber seedlings inoculated with strain 62 and showing disease symptoms on cotyledons only. This implied that water stress was not limited to symptomatic cotyledons but extended to the whole seedling.     

Use of NMR for measurement of total body water and estimation of body fat

A nuclear magnetic resonance (NMR) method is described for quantitatively measuring total body water (TBW) and for estimating the fat content of baboons. The hydrogen associated with water was measured as the amplitude of the free-induction decay voltage following a series of 90 degree radio frequency pulses at the Lamour frequency for hydrogen with a pulse length of 14 microseconds and a peak measuring time of 50 microseconds. TBW was calculated by multiplying the peak amplitude (volts) by the experimentally determined constant for a water standard (g water/V). This NMR method yielded TBW contents similar to those obtained in the same baboons by direct gravimetric procedures. In contrast, the widely used 3H2O-dilution method usually and variably overestimated body water. By providing an accurate measure of body water, this NMR procedure provides a rapid, noninvasive, reasonably accurate way of estimating body fat content.     

黄瓜叶片光合电子传递对水分胁迫的响应

黄瓜叶片在水分胁迫下叶片相对含水量减少,类囊体室温吸收光谱的吸收峰降低,同时其NADP光还原活性、Ca^2 -ATPase活性也相应降低,全链电子传递明显受阻。类囊体膜蛋白电泳分析结果显示：类囊体膜色素蛋白复合体含量有不同程度的降低,其中PSⅡ色素蛋白复合体含量下降较多,试验结果表明水分胁迫通过限制光能的吸收,传递双及转换效率,抑制了光合电子传递过程。     

Enantioselective determination of triazole fungice tetraconazole by chiral high-performance liquid chromatography and its application to pharmacokinetic study in cucumber, muskmelon, and soils

A simple chiral high-performance liquid chromatography method with diode array detector was developed and validated for stereoselective determination of tetraconazole enantiomers in cucumber, muskmelon, and soils. Good separation was achieved at 20°C using cellulose tris-(4-methylbenzoate) as chiral stationary phase, a mixture of n-hexane and ethanol (90:10) as mobile phase at a flow rate of 0.8 ml/min. The assay method was linear over a range of concentrations (0.5-50 μg/ml) and the mean recoveries in all samples were more than 85% for the two enantiomers. The limits of detection for both enantiomers in plant and soil samples were 0.06 and 0.12 μg/g, respectively. Then, the proposed method was successfully applied to the study of enantioselective degradation of rac-tetraconazole in cucumber, muskmelon, and soils. The results showed that the degradation of two enantiomers of tetraconazole followed first-order kinetics and significantly stereoselective behavior was observed in cucumber, muskmelon, and Beijing soil. The preferential absorption and degradation of (-)-S-tetraconzole resulted in an enrichment of the (+)-R-tetraconazole residue in plant samples, whereas the (+)-R-tetraconazole showed a faster degradation in Beijing soil and the stereoselectivity might be caused by microorganisms. No stereoselective degradation was observed in Heilongjiang soil.     

Flow mixing enhancement from balloon pulsations in an intravenous oxygenator

Computational investigations of flow mixing and oxygen transfer characteristics in an intravenous membrane oxygenator (IMO) are performed by direct numerical simulations of the conservation of mass, momentum, and species equations. Three-dimensional computational models are developed to investigate flow-mixing and oxygen-transfer characteristics for stationary and pulsating balloons, using the spectral element method. For a stationary balloon, the effect of the fiber placement within the fiber bundle and the number of fiber rings is investigated. In a pulsating balloon, the flow mixing characteristics are determined and the oxygen transfer rate is evaluated. For a stationary balloon, numerical simulations show two well-defined flow patterns that depend on the region of the IMO device. Successive increases of the Reynolds number raise the longitudinal velocity without creating secondary flow. This characteristic is not affected by staggered or non-staggered fiber placement within the fiber bundle. For a pulsating balloon, the flow mixing is enhanced by generating a three-dimensional time-dependent flow characterized by oscillatory radial, pulsatile longitudinal, and both oscillatory and random tangential velocities. This three-dimensional flow increases the flow mixing due to an active time-dependent secondary flow, particularly around the fibers. Analytical models show the fiber bundle placement effect on the pressure gradient and flow pattern. The oxygen transport from the fiber surface to the mean flow is due to a dominant radial diffusion mechanism, for the stationary balloon. The oxygen transfer rate reaches an asymptotic behavior at relatively low Reynolds numbers. For a pulsating balloon, the time-dependent oxygen-concentration field resembles the oscillatory and wavy nature of the time-dependent flow. Sherwood number evaluations demonstrate that balloon pulsations enhance the oxygen transfer rate, even for smaller flow rates.     

Flow analysis by phase modulation of NMR signals

Time modulated field-gradient-sequences able to selectively phase encode spatial location or flow velocity in NMR signals are described. The specific phase modulation discriminates between stationary and mobile protons. The method is presented in the one dimensional case and has the following advantages: it is sensitive to flow direction and unaffected by the relaxation times.     

消毒、伴生和微生物菌对连作基质性质及黄瓜生长发育的影响

以连续种植2茬的连作基质为对象,采用室内盆栽试验,设置连作基质太阳能消毒、热水消毒、多菌灵消毒、大蒜伴生栽培和施入微生物菌处理以及新基质对照,在种植黄瓜后观测基质理化性质、养分含量、酚类物质含量、相关酶活性的变化以及黄瓜结果期生长发育、光合作用和果实产量的变化,探究不同处理对连作基质栽培黄瓜生长发育影响的机制,以期在生产上为废旧基质重复利用提供新的思路和方法。结果表明:在连作基质栽培黄瓜后,太阳能消毒和热水消毒处理均能显著提高连作基质的pH,同时降低电导率;施入微生物菌处理的基质EC值、速效氮、速效磷、速效钾和总酚含量均显著低于连作基质,而其基质磷酸酶活性和过氧化氢酶活性显著高于连作基质。热水消毒还显著降低连作基质的总酚含量和复合态酚含量。黄瓜生长至结果期,太阳能消毒、热水消毒、多菌灵消毒、大蒜伴生栽培和施入微生物菌均显著增强黄瓜的根系活力和株高;其中热水消毒和施入微生物菌处理的产量增加最为显著,分别比连作基质栽培提高了16.56%和18.40%。可见,连作基质进行热水消毒处理和施入微生物菌处理均能降低基质盐分积累,减少总酚含量,进而显著提高黄瓜的根系活力和光合作用,从而增加产量。     

In situ plant water balance studies using a portable NMR spectrometer

A portable ¹H NMR spectrometer has been applied to whole plantsin situ, i.e. in climate rooms and in a greenhouse, to studyplant water relations under these conditions. The spectrometerconsists of a 30 kg permanent magnet system of 0.235 T, modifiedBruker Minispec electronics, and a standard XT pc for spectrometercontrol. Unattended, the system automatically measures the xylemsap stream and tissue water content in a well-defined sectionof the plant stem. Because of the small size of the magnet thesemeasurements can be made at different positions on the plantstem or on different plants in succession. Magnetic field driftdue to the varying climates that occur in greenhouses was correctedby field locking. Results are presented for a single plant ina climate room, demonstrating a method for the study of plantwater hydraulics. In addition, a single plant in a greenhousecrop was measured, and the NMR results were compared with thewater uptake and transpiration rates under (natural) variationof light intensity and relative humidity, demonstrating thereliability of the portable NMR under realistic greenhouse conditions.Finally, the application for the measurement of the root wateruptake efficiency is demonstrated for a number of grafted cucumberplants under constant climatic conditions in a phytotron. Key words: Portable NMR system, plant water balance sensor, greenhouse applications, flow, water content     

Gating of aquaporins by low temperature in roots of chilling-sensitive cucumber and chilling-tolerant figleaf gourd

Effects of low temperature (8 degrees C) on the hydraulic conductivity of young roots of a chilling-sensitive (cucumber, Cucumis sativus L.) and a chilling-resistant (figleaf gourd, Cucurbita ficifolia Bouche) crop have been measured at the levels of whole root systems (root hydraulic conductivity, Lp(r)) and of individual cortical cells (cell hydraulic conductivity, Lp). Exposure of roots to low temperature (LRT) for up to 6 d caused a stronger suberization of the endodermis in cucumber compared with figleaf gourd, but no development of exodermal Casparian bands in either species. Changes in anatomy after 6 d of LRT treatment corresponded with a reduction in hydrostatic root Lp(r) of cucumber roots by a factor of 24, and by a factor of 2 in figleaf gourd. In figleaf gourd, there was a reduction only in hydrostatic Lp(r) but not in osmotic Lp(r) suggesting that the activity of water channels was not much affected by LRT treatment in this species. Changes in cell Lp in response to chilling and recovery were similar to the root levels, although they were more intense at the root level. Activation energies (E(a)) and Q10 of water flow as measured at the cell level were high in cucumber (E(a)=109+/-13 kJ mol(-1); Q(10)=4.8+/-0.7; n=6-10 cells), but small in figleaf gourd (E(a)=11+/-2 kJ mol(-1); Q10=1.2+/-0.1; n=6-10 cells). Roots of figleaf gourd recovered better from LRT treatment than those of cucumber. In figleaf gourd, recovery (at both the root and cell level) often resulted in Lp and Lp(r) values which were even bigger than the original, i.e. there was an overshoot in hydraulic conductivity. These effects were larger for osmotic (representing the cell-to-cell passage of water) than for hydrostatic Lp(r). After a short-term (1 d) exposure to 8 degrees C followed by 1 d at 20 degrees C, hydrostatic Lp(r) of cucumber nearly recovered and that of figleaf gourd still remained higher due to the overshoot. By contrast, osmotic Lp(r) and cell Lp in both species remained high by a factor of 3 compared with the control, possibly due to an increased activity of water channels. After preconditioning of roots at LRT, increased hydraulic conductivity was completely inhibited by HgCl2 at both the root and cell levels. Different from figleaf gourd, recovery from chilling was not complete in cucumber after longer exposure to LRT. It is concluded that at LRT, both changes in the activity of aquaporins (AQPs) and alterations of root anatomy determine the water uptake in both species. The high temperature dependence of cell Lp in cucumber suggests conformational changes of AQPs during LRT treatment which result in channel closure and in a strong gating of AQP activity by low temperature. This mechanism is thought to be different from that in figleaf gourd where AQPs reacted in the conventional way, i.e. low temperature affected the mobility of water molecules in AQPs rather than their open/closed state, and Q(10) was low.     

土壤水分对温室嫁接和非嫁接黄瓜生长与生理特性的影响

观测了黄瓜生长动态与生理特性对土壤水分状况的反应。结果表明,日光温室嫁接与未嫁接黄瓜分别在土壤相对含水量70％和80％时株高,叶面积和产量增长最快,土壤水分过高与过低均不利于黄瓜的生长;随土壤含水量降低;叶片光合速率,气孔导度,水势与根系活力的下降。     

Measurement of Lipid Accumulation in Chlorella vulgaris via Flow Cytometry and Liquid-State 1H NMR Spectroscopy for Development of an NMR-Traceable Flow Cytometry Protocol

In this study, we cultured Chlorella vulgaris cells with a range of lipid contents, induced via nitrogen starvation, and characterized them via flow cytometry, with BODIPY 505/515 as a fluorescent lipid label, and liquid-state ¹H NMR spectroscopy. In doing so, we demonstrate the utility of calibrating flow cytometric measurements of algal lipid content using triacylglyceride (TAG, also known as triacylglycerol or triglyceride) content per cell as measured via quantitative ¹H NMR. Ensemble-averaged fluorescence of BODIPY-labeled cells was highly correlated with average TAG content per cell measured by bulk NMR, with a linear regression yielding a linear fit with r ² = 0.9974. This correlation compares favorably to previous calibrations of flow cytometry protocols to lipid content measured via extraction, and calibration by NMR avoids the time and complexity that is generally required for lipid quantitation via extraction. Flow cytometry calibrated to a direct measurement of TAG content can be used to investigate the distribution of lipid contents for cells within a culture. Our flow cytometry measurements showed that Chlorella vulgaris cells subjected to nitrogen limitation exhibited higher mean lipid content but a wider distribution of lipid content that overlapped the relatively narrow distribution of lipid content for replete cells, suggesting that nitrogen limitation induces lipid accumulation in only a subset of cells. Calibration of flow cytometry protocols using direct in situ measurement of TAG content via NMR will facilitate rapid development of more precise flow cytometry protocols, enabling investigation of algal lipid accumulation for development of more productive algal biofuel feedstocks and cultivation protocols.     

Disappearance of desiccation tolerance of imbibed crop seeds is not associated with the decline of oligosaccharides

The relationship between sugar content and loss of desiccation tolerance of hydrated crop seeds (tomato, okra, snow pea, mung bean, and cucumber) was evaluated by imbibing seeds with or without ABA, followed by dehydration and germination. During the process of hydration, but before the seeds lost desiccation tolerance, monosaccharide content increased only slightly, sucrose increased in snow peas, mung bean and cucumber, but maintained its original level in other species and the oligosaccharides declined dramatically. At the time of losing desiccation tolerance, the sucrose content of imbibed seeds was 2-3 times higher than the original level in most species. Positive significant correlation coefficients (r) were found in many, but not all crop seeds between desiccation tolerance and the oligosaccharide mass, or oligo/sucrose ratio. The ratio of oligo/sucrose in intact seeds at the time of losing desiccation tolerance, however, was not a fixed value and varied among species. Oligosaccharides declined significantly in different seed parts of imbibed cucumber seeds while sucrose increased to a higher level in the radicle than in the hypocotyl. Radicles were far more sensitive to desiccation than hypocotyls. The same observation was found for cucumber seeds imbibed in 100 M ABA, yet desiccation tolerance was largely maintained in hypocotyls and cotyledons. It is concluded that sucrose and oligosaccharides are not the determinants of the loss of desiccation tolerance in hydrated seeds.Imbibed seeds did not show any differences between seed parts in their ability to resynthesize sugars during the process of slow dehydration. Differences in sensitivity to desiccation among seed parts were not due to differences in the initial water content or to the rate of water content increase among seed parts. Physiological regulation of the loss of desiccation tolerance in crop seeds during hydration is discussed.