Water Deficit and Salt Stress Diagnosis Through LED Induced Chlorophyll Fluorescence Analysis in Jatropha curcas L.

LED induced chlorophyll fluorescence analysis is employed to investigate the effect of water deficit and salt stress upon the growth process of Jatropha curcas L.. Red(Fr) and far-red(FFr) chlorophyll fluorescence around 685 nm and 735 nm, respectively, were observed and examined as a function of the stress intensity(salt concentration and water deficit). The fluorescence ratio Fr/FFr which is a valuable nondestructive and nonintrusive indicator of the chlorophyll content of leaves was exploited to monitor the jatropha plants under stress. The data indicated that salinity plays a minor role in the chlorophyll concentration of leaves for NaCl concentrations in the 25 to 200 mM range. The fluorescence ratio also permitted the detection of damage caused by water deficit in the early stages of the plants growing process. A significant variation of the Fr/FFr ratio was observed in the first 10 days of the experiment, and before signs of visual stress became apparent. The results suggest that the Fr/FFr ratio is an early-warning indicator of water deficit stress     

叶绿素荧光及其在水分胁迫监测中的研究进展

水分胁迫是一种常见的植被胁迫状态,能够减小细胞水势,引起气孔关闭,破坏光合进程。叶绿素荧光是植物光合作用的探针,可以很好的反应植物生理特征,能够快速、灵敏、无损伤和深层次监测水分胁迫状态。由于叶绿素荧光的影响因素较多,使得水分胁迫与叶绿素荧光的关系比较复杂。从光合作用角度阐述了叶绿素荧光的产生原理,分析了叶绿素、叶片结构、冠层结构和环境因子等对叶绿素荧光的影响,总结了叶绿素荧光在水分胁迫监测中的应用;针对叶绿素荧光复杂的生理基础、影响因素及其在水分胁迫监测中的研究进展,提出了一些科学问题和未来的研究方向,为叶绿素荧光的进一步深入研究提供参考,为叶绿素荧光遥感的进一步应用提供理论支持,为陆地生态系统的准确评估提供借鉴。     

Water stress assessment of cork oak leaves and maritime pine needles based on LIF spectra

The aim of the present work was to develop a method for the remote assessment of the impact of fire and drought stress on Mediterranean forest species such as the cork oak (Quercus suber) and maritime pine (Pinus pinaster). The proposed method is based on laser induced fluorescence (LIF): chlorophyll fluorescence is remotely excited by frequency-doubled YAG:Nd laser radiation pulses and collected and analyzed using a telescope and a gated high sensitivity spectrometer. The plant health criterion used is based on the I ₆₈₅/I ₇₄₀ ratio value, calculated from the fluorescence spectra. The method was benchmarked by comparing the results achieved with those obtained by conventional, continuous excitation fluorometric method and water loss gravimetric measurements. The results obtained with both methods show a strong correlation between them and with the weight-loss measurements, showing that the proposed method is suitable for fire and drought impact assessment on these two species.     

农田土壤水分含量的激光诱导荧光光谱表征

通过实验探讨了植物的激光诱导叶绿素荧光光谱与土壤水分间的关系。实验利用Y-形光纤探头获取了植物在450 nm LED光源照射下所发出的叶绿素荧光光谱,同时利用TDR传感器测量土壤湿度。实验以水稻为研究对象,研究了在持续水分胁迫和间歇水分胁迫下,叶绿素荧光光谱743 nm附近波峰的变化,结果发现,波峰强度与土壤水分含量具有相关性。最后,利用Lorentzian方程,将持续水分胁迫下的土壤含水量与叶绿素荧光强度进行建模,发现所建的模型具有较高的决定系数,说明该方法可以应用于农业生产中对土壤水分的测量。     

Stress-Induced Alteration of Chlorophyll Fluorescence Polarization and Spectrum in Leaves of <Emphasis Type="Italic">Alocasia macrorrhiza</Emphasis> L. Schott

The value of intrinsic chlorophyll fluorescence polarization, and the intensity in emission spectrum were investigated in leaf segments of Alocasia macrorrhiza under several stress conditions including different temperatures (25–50°C), various concentrations of NaCl (0–250 mM), methyl viologen (MV, 0–25 μM), SDS (0–1.0%) and NaHSO₃ (0–80 μM). Fluorescence emission spectrum of leaves at wavelength regions of 500–800 nm was monitored by excitation at 436 nm. The value of fluorescence polarization (P value), as result of energy transfer and mutual orientation between chlorophyll molecules, was determined by excitation at 436 nm and emission at 685 nm. The results showed that elevated temperature and concentrations of salt (NaCl), photooxidant (MV), surfactant (SDS) and simulated SO₂ (NaHSO₃) treatments all induced a reduction of fluorescence polarization to various degrees. However, alteration of the fluorescence spectrum and emission intensity of F₆₈₅ and F₇₃₁ depended on the individual treatment. Increase in temperature and concentration of NaHSO₃ enhanced fluorescence intensity mainly at F₆₈₅, while an increase in MV concentration led to a decrease at both F₆₈₅ and F₇₃₁. On the contrary, NaCl and SDS did not cause remarkable change in fluorescence spectrum. Among different treatments, the negative correlation between polarization and fluorescence intensity was found with NaHSO₃ treatments only. We concluded that P value being measured with intrinsic chlorophyll fluorescence as probe in leaves is a susceptible indicator responding to changes in environmental conditions. The alteration of P value and fluorescence intensity might not always be shown a functional relation pattern. The possible reasons of differed response to various treatments were discussed.     

Estimation of leaf chlorophyll content and density in Populus euphratica based on hyperspectral characteristic variables

Abstract

Chlorophylls are the most important pigments for photosynthesis and are related to the phenology and health status of plants. Monitoring the chlorophyll content and density of Populus euphratica leaves via remote sensing technology has great significance for restoring and reconstructing damaged desert ecosystems. In this study, Populus euphratica leaves were collected in the public welfare forest reserve of Awati county, Xinjiang, China. Spectral variables and the chlorophyll content and density of leaves were obtained, simple linear regression and multiple linear regression models were built to predict leaf chlorophyll content and density. The results showed that partial least-squares regression models are the most appropriate for chlorophyll content estimation and its determination coefficient and root mean square error were 0.630?～?0.745 and 0.031?～?0.240, respectively. The standard deviation to root mean square error ratio was greater than 1.8 and the ratio of the standard error of the laboratory to the standard error of prediction was close to 1 for the partial least-squares regression models. These results confirmed that chlorophyll content can be accurately predicted and the proposed partial least-squares regression models were adequate for estimating Populus euphratica chlorophyll content rapidly and nondestructively, representing an alternative to conventional measurement methods. The rapid monitoring of chlorophyll content can reflect the health condition of Populus euphratica in a timely manner and provide technical support for ecological reconstruction in arid areas.     

水稻叶绿素浓度对荧光光谱的影响

为了加强对水稻生长状况的监测,指导水稻的田间施肥,提高施肥利用效率,以增加农作物产量、提高粮食品质,实验室搭建了基于激光诱导荧光技术的荧光探测系统,以研究水稻叶片的叶绿素含量与荧光峰值比之间的相关性。文中测量样本为水稻分蘖期和拔节期的倒二叶,栽培地区位于中国江汉平原。文中先采用凯氏定氮法和相应公式结合测得水稻叶片的叶绿素含量(mg·g-1),再用搭建的荧光探测系统采集了水稻叶片不同叶绿素含量的荧光光谱(激发波长为355 nm)。获得了水稻叶片在不同叶绿素含量下的荧光光谱数据库,定量分析了荧光峰值比F₇₄₀/F₆₈₅(荧光谱峰740 nm、685 nm处的荧光强度比)与叶绿素含量的相关性,发现叶绿素含量的变化对荧光光谱特性影响明显。由实验数据分析可知荧光参数中的峰值比(F₇₄₀/F₆₈₅)与叶绿素含量呈现很好的线性正相关,相关系数(R2)在水稻的分蘖期和拔节期分别达到了0.901 3和0.912 5。实验分析结果表明诱导荧光光谱技术具有方便、快捷、无损等优点,在农作物生长状况的遥感定量监测等方面也具有一定发展潜力。     

不同水分处理下草莓叶绿素荧光参数的变化

以草莓为实验材料,研究不同水分处理对叶片叶绿素荧光参数的影响.研究表明,随着施水量的增加,草莓叶片的叶绿素荧光参数存在着明显的差异,过多或过少的水分均降低了叶片的PSⅡ活性.总体看来,保持土壤含水量为田间最大持水量的80%(T8)时的最大荧光(Fm)、PS Ⅱ最大光化学效率(Fv/Fm)、PS Ⅱ实际光化学效率(Yie...     

饱和激发下叶绿素荧光非线性变化及影响分析

叶绿素浓度是海洋初级生产力的重要指标之一,激光诱导荧光技术可以实现海水叶绿素浓度的快速测量。测量叶绿素浓度的传统激光诱导荧光原理,是利用叶绿素荧光与水体Raman散射的强度比值（I_F/R）进行反演,即叶绿素浓度n_chl＝CI_F/R,其中C为系统常量。这是依据叶绿素荧光685 nm、水体Raman散射强度都与激发光强呈线性关系。然而,该理论并没有考虑诱导荧光饱和现象的存在。当诱导激光强度达到一定程度后,685 nm荧光强度随激发光强非线性变化。另外,值得注意的是,水体Raman散射并不存在信号饱和现象。为了探讨饱和激发造成荧光非线性变化的影响,在激光诱导荧光技术测量叶绿素浓度的实验中,设计两种测量方案,即：不同激光功率诱导单一浓度样本的荧光测量,和固定激光功率时不同浓度样本的荧光测量。实验中利用Nd∶YAG三倍频激光355 nm激发获得叶绿素溶液的404 nm处 Raman散射和685 nm荧光。实验结果分为2部分进行讨论：（1）为了分析饱和激发造成荧光变化的非线性特性,通过调节激发光功率测量溶液的受激发射光谱,发现水体Raman散射强度与激发光强呈线性关系,而685 nm荧光强度出现饱和激发下的非线性变化。而且,随叶绿素浓度的增加,685 nm荧光的非线性趋势更为明显,Raman散射强度与激发光强的线性关系中斜率变小。数据分析表明,685 nm荧光数据拟合的4阶多项式和Raman散射效率值,可以定性地表征685 nm荧光的饱和程度。（2）考虑实际海洋激光雷达探测叶绿素浓度应用中存在饱和激发荧光非线性现象,为了分析荧光非线性对传统叶绿素浓度反演理论适用性的影响,在固定激发光强情况下对不同浓度叶绿素溶液的发射光谱进行测量。将激发光功率调节至52.00,80.70,132.10和197.30 mW·cm-2,获取相应激发光强下685 nm荧光与水体Raman散射的强度比值和叶绿素浓度之间的关系。实验表明,激发光强不变的情况下,685 nm荧光与水体Raman散射的强度比值,与叶绿素浓度仍满足线性关系。但是,在较高光强激发时,饱和激发造成的叶绿素荧光非线性变化,导致利用传统激光诱导荧光理论反演的叶绿素浓度值偏小。因此,需要对饱和激发下荧光非线性的影响进行修正,其关系为I_F/R=n_chl/C+C_F,修正值C_F不可忽略。另外,值得一提的是,修正关系中系统常量C随激发光强增加而增大。研究表明,饱和激发造成的荧光非线性,会对激光诱导荧光技术测量叶绿素浓度产生影响,但由于造成荧光非线性因素的复杂性,仅通过荧光数据拟合获得的多项式,无法定量说明其影响权重。然而,当激发光强不变时,可以实验测量获得基于激光诱导荧光原理的修正关系,从而准确反演叶绿素浓度。     

基于叶绿素荧光诱导动力学曲线的光合作用参数反演算法研究

快相叶绿素荧光诱导曲线中蕴含着丰富光合作用信息。该信息可以反映出植物的生存状态、病理以及受到胁迫时的生理变化趋势等多种信息。通过采集藻类荧光及诱导光信号, 拟合快相叶绿素荧光动力学曲线。基于最小二乘拟合方法,提出自适应最小误差逼近的方法对快相叶绿素荧光动力学曲线进行多元非线性回归拟合,实现F_o（固定荧光）、F_m（最大荧光产率）、σ_PSII（PSII的功能吸收截面）等细节参数的反演。实现了蛋白核小球藻光合作用参数反演,并实验了在Cu2+胁迫环境下,蛋白核小球藻的生理变化趋势。     

Performance Check of Vegetation Fluorescence Imaging Lidar through In Vivo and Remote Estimation of Chlorophyll Concentration Inside Plant Leaves

Laser-induced fluorescence imaging lidar was developed for in vivo plant/vegetation monitoring. Fluorescences of poplar tree leaves growing naturally at a distance of 60 m from the lidar were successfully detected as two spectral images at wavelengths of 685 nm and 740 nm. By comparing chlorophyll concentration quantified with high performance liquid chromatography, it was confirmed that the intensity ratio of the two wavelengths was converted into the chlorophyll concentration inside the leaves. The intensity ratio of the images reflected the status of the poplar tree in the process of senescence so well that it was possible to assess the living status of the tree as a numerical value. The performance characteristics and the potential of the laser-induced fluorescence imaging lidar for monitoring the physiological activities of plants and vegetation are described.     

A New Dual-peak Fluorescent Probe for Water Content Detection Made From Taxus

In this paper, the leaves of Taxus were used as the sole carbon source, and two kinds of carbon dots blue and red, with different properties, were synthesized by the hydrothermal method under different conditions. The red carbon dots were quenched in the water, and the blue carbon dots had stable fluorescence properties in water environment. The bimodal fluorescence probe formed by mixing could accurately and stably measure the water content in ethanol, which was in the range of 82.5%-100%, is highly correlated with the fluorescence intensity ratio (I₄₈₁/I₆₇₈) of mixed carbon dots under 390 nm excitation light, with R²?=?0.995 and the detection limit as low as 0.31%. The experimental materials are environmentally friendly, low in cost, and simple to operate, as well as the water content measured by proportional fluorescence has high accuracy, which provides a new method for measuring moisture in ethanol.

     

Fluorescence of bean leaves grown under low-light conditions

The patterns of changes of the spectral shapes of the fluorescence of bean leaves have been identified under low-light conditions of plant cultivation and decrease in chlorophyll concentration in the leaves. Under these experimental conditions, the ratio of peak intensities of fluorescence in the red spectral region, ω = F ₇₄₀/F ₇₆₀, which was recorded after the end of the induction period, was proportional to the chlorophyll content per 1 g of the wet weight of a leaf.     

Measurement of Cell Volume Changes by Fluorescence Self-Quenching

At high concentrations, certain fluorophores undergo self-quenching, i.e., fluorescence intensity decreases with increasing fluorophore concentration. Accordingly, the self-quenching properties can be used for measuring water volume changes in lipid vesicles. In cells, quantitative determination of water transport using fluorescence self-quenching has been complicated by the requirement of relatively high (mM) and often toxic loading concentrations. Here we report a simple method that uses low (M) loading concentrations of calcein-acetoxymethyl ester (calcein-AM) to obtain intracellular concentrations of the fluorophore calcein suitable for measurement of changes in cell water volume by self-quenching. The relationship between calcein fluorescence intensity, when excited at 490 nm (its excitation maximum), and calcein concentration was investigated in vitro and in various cultured cell types. The relationship was bell-shaped, with the negative slope in the concentration range where the fluorophore undergoes fluorescence self-quenching. In cultured retinal pigment epithelial cells, calcein fluorescence and extracellular osmolarity were linearly related. A 25-mOsm hypertonic challenge corresponded to a decrease in calcein fluorescence with high signal-to-noise ratio (>15). Similar results were obtained with the fluorophore BCECF when excited at its isosbestic wavelength (436 nm). The present results demonstrate the usefulness of fluorescence self-quenching to measure rapid changes in cell water volume.     

Efficiency of fluorescence excitation in chlorophyll <Emphasis Type="Italic">a</Emphasis> by the second and third harmonics of emission from an Nd:YAG laser

We have studied the efficiency of fluorescence excitation in chlorophyll a by the second harmonic (532 nm) and the third harmonic (355 nm) of an Nd:YAG laser for sea water samples with phytoplankton taken during different algal blooms and for extracts of higher plants diluted with sea water. We have shown that when using the second harmonic, the efficiency of fluorescence excitation in chlorophyll a is more than twice as high as for excitation by the third harmonic. We discuss the advisability of using the second or third harmonics of the emission from an Nd:YAG laser when studying the biooptical parameters of sea water. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 75, No. 2, pp. 224–227, March–April, 2008.     

Analysis of Chlorophyll Fluorescence Spectra in Some Tropical Plants

Laser-induced chlorophyll fluorescence (LICF) emission spectra of leaves of some tropical plants were measured using a compact fiber-optic fluorosensor with a continuous-wave blue diode laser as exciting source and an integrated digital spectrometer. Different chlorophyll-fluorescence signatures of light-green, fully-green, and yellow leaves were monitored at room temperature. Deconvolution procedure was used to determine fluorescence band position and width. Calibration of the fluorescence ratio F690/F730 relative to the 404 nm excitation is done from the curve-fitted parameter. An erratum to this article can be found at     

Spectral changes in the fluorescence of chlorophyll during photosynthesis induction

With the help of a light-emitting diode with a radiation maximum at 407 nm and an S-2000 UV-VIS spectrometer connected with a computer, the spectral changes in the fluorescence of chlorophyll from acacia leaves (Acacia sp.) preliminarily subjected to a dark adaptation are studied. It is found that, in the slow induction phase, the Kautsky effect manifests itself in the “compression” of the intensity of the chlorophyll fluorescence spectrum, with this spectrum exponentially approaching the steady-state shape with a time constant of 10–20 s. Once the steady-state fluorescence spectrum of chlorophyll was established, the amount of energy delivered to the upper singlet level is one and a half times greater than at 735 nm. The ratio between the energies spent for photochemical processes of photosynthesis and for fluorescence depends on the wavelength and the instant of time of the induction period. In the steady-state state of the chlorophyll fluorescence, the values of this ratio at 685 and 735 nm are equal to 5.9 and 3.4, respectively.     

激光诱导植物荧光寿命校正技术及其特性分析

激光诱导植物荧光寿命测量法是在植物荧光光谱分析法基础上开发的一种评估植物生长状况及环境监测的新技术。根据植物叶绿素荧光信号的物理特性,利用信息仿真技术开发了一种叶绿素荧光寿命校正方法,可提高植物叶绿素荧光寿命的测量精度。利用激光诱导叶绿素荧光寿命测量系统分别测得叶绿素荧光及其背景信号,先用解卷积法叶绿素荧光信号中分离出荧光衰减函数,可获取荧光寿命估计值。再结合叶绿素荧光寿命校正技术就能反演得到高精度的植物荧光寿命。仿真与实验结果表明：该方法可实现高精度的植物荧光寿命实时监测;并对不同含量的叶绿素提取液进行了测试,构建了植物荧光寿命与叶绿素含量的对应关系模型。未来该技术可用于遥感监测海洋、湖泊、河流中藻类植物的生物含量。     

铜胁迫下玉米叶片反射光谱的红边位置变化及其与叶绿素的关系

采用5级梯度铜胁迫砂培试验,通过测定叶片反射光谱曲线和叶绿素含量,研究铜胁迫对玉米叶片反射光谱与叶绿素含量的影响。研究表明：玉米叶片反射光谱的红边位置与铜胁迫浓度显著相关(R2≥0.5755),且存在明显的“红边蓝移”现象,即叶片光谱红边位置向短波方向移动。随着铜胁迫浓度的升高或胁迫时间延长,红边蓝移程度增加。不同胁迫浓度之间,叶绿素a、叶绿素b含量及二者比值 (Chla/Chlb)均差异显著(p分别为0.002,0.007和0.001)。叶绿素a、b比值(Chla/Chlb)与培养液中铜浓度呈显著负相关(R=-0.898);Chla/Chlb与平均红边波长显著正相关(R=0.814)。这表明随着铜胁迫浓度升高,Chla/Chlb降低,叶绿素b相对于叶绿素a升高,叶片反射光谱的红边位置蓝移。铜胁迫改变了玉米叶片中Chla和Chlb含量和比值,由此导致的色素吸收光谱变化,是铜胁迫导致叶片反射光谱红边蓝移的可能的生理学成因。     

Determining the Canopy Water Stress for Spring Wheat Using Canopy Hyperspectral Reflectance Data in Loess Plateau Semiarid Regions

Crop water stress significantly reduces crop yield. Several studies have employed optical and remote-sensing methods to obtain nondamage monitoring crop water content to understand the agriculture drought process. In this paper, the spectral information (i.e., the canopy absorption feature at the 350–2500 nm band range) from the field experiments was used to estimate and identify the canopy water stress. Five different levels of water treatments exist in the spring wheat field in the semiarid regions of Loess Plateau, Northwest China. The hyperspectral reflectance, soil moisture content, soil water potential, canopy water content, amount of chlorophyll, leaf area index, and environmental parameters were measured. The relationship between canopy reflectance and canopy water content was analyzed at different water stress levels. In addition, various spectral indices were tested by measurements. Results showed that a high correlation exists in semiarid water index-1, semiarid water index-2, and red-edge normalized difference vegetation index, thus denoting that these indices can indicate water stress effectively. We can conclude that canopy reflectance can identify crop water stress and can be used to develop a certain index for monitoring agriculture drought.