Development of a digital image analysis method for real-time estimation of chlorophyll content in micropropagated potato plants

The present work describes a digital image analysis method based on leaf color analysis to estimate chlorophyll content of leaves of micropropagated potato plantlets. For estimation of chlorophyll content, a simple leaf digital analysis procedure using a simple digital still camera was applied in parallel to a SPAD chlorophyll content meter. RGB features were extracted from the image and correlated with the SPAD values. None of the mean brightness parameters (RGB) were correlated with the actual chlorophyll content following simple correlation studies. However, a correlation between the chromaticity co-ordinates ‘r’, ‘b’ and chlorophyll content was observed, while co-ordinate ‘g’ was not significantly correlated with chlorophyll content. Linear regression and artificial neural networks (ANN) were applied for correlating the mean brightness (RGB) and mean brightness ratio (rgb) features to chlorophyll content of plantlet leaves determined through a SPAD meter. The chlorophyll content as determined by the SPAD meter was significantly correlated (RMSE = 3.97 and 3.59, respectively, for linear and ANN models) to the rgb values of leaf image analysis. Both the models indicate successful prediction of chlorophyll content of leaves of micropropagated plants with high correlation. The developed RGB-based digital image analysis has the advantage over conventional subjective methods for being objective, fast, non-invasive, and inexpensive. The system could be utilized for real-time estimation of chlorophyll content and subsequent analysis of photosynthetic and hyperhydric status of the micropropagated plants for better ex vitro survival.     

Simultaneous estimation of chlorophyll a and lipid contents in microalgae by three-color analysis

A method of rapid determination of chlorophyll a and lipid contents of microalgae based on colorimetric analysis of the digital images of the microalgae is proposed. The color variation of microalgae during cultivation is evaluated by the brightness of the three primary colors (red, green, and blue). The brightness values of the three primary colors are modeled as two linear correlation functions (RGB model) for microalgal chlorophyll a and lipid contents, respectively. The chlorophyll a and lipid contents predicted by the proposed model are compared with that determined by the standard methods. The good agreement of the model predictions with experimental results is demonstrated with a squared correlation coefficient (R(2)) of 0.99 for chlorophyll a and lipid. The reliability of the RGB model was verified in real cultivations of the microalgae in a photobioreactor. Growth dynamics, contents of chlorophyll a and lipid corresponded very well with previously reported studies.     

枫香秋季变色期叶色变化及其生理基础

为探究秋季枫叶呈色的关键生理因素,该文以转色期叶色为绿色、黄色和红色的枫香单株为试材,研究了L^*、a^*、b^*值变化与叶片色素、可溶性糖及可溶性蛋白质含量变化的相关性。结果表明:(1)在变色期,3种色彩枫香叶片叶绿素a、叶绿素b、总叶绿素和类胡萝卜素均大量降解,花色素苷不同程度积累。(2)绿色叶单株叶绿素和类胡萝卜素始终保持较高含量,花色素苷含量上升4.2倍,叶片内色素含量比值始终保持稳定; 黄色叶单株叶绿素和类胡萝卜素含量最低,花色素苷含量上升4.4倍,b^*值与叶绿素含量极显著负相关,与类胡萝卜素含量显著负相关,与花色素苷/类胡萝卜素含量比值极显著正相关; 红色叶单株叶绿素和类胡萝卜素含量略高于黄色叶单株,花色素苷含量上升27.2倍,a^*值与叶绿素含量、类胡萝卜素含量极显著负相关,与花色素苷含量显著正相关,与色素含量比值无显著相关性。(3)红色叶单株具有较高的可溶性糖含量和可溶性蛋白质含量。因此,在枫香叶片变色期,保持较高的叶绿素和类胡萝卜素含量,维持色素含量比值稳定使叶片呈现绿色; 叶绿素和类胡萝卜素的大量降解,以及花色素苷/类胡萝卜素含量比值的升高使叶片呈现黄色; 叶绿素的降解和花色素苷的大量合成使叶片呈现红色。     

Intelligent image analysis (IIA) using artificial neural network (ANN) for non-invasive estimation of chlorophyll content in micropropagated plants of potato

An artificial neural network (ANN) was used to analyze photometric features extracted from the digitized images of leaves from in vitro-regenerated potato plants for non-invasive estimation of chlorophyll content. A MATLAB®-based, feed-forward, backpropagation-type network was developed for an input layer (three input elements), with one hidden layer (one node) and one output layer representing the predicted chlorophyll content. A significant influence of training function during optimization of ANN modeling was observed. Among the 11 training functions tested, “trainlm” was found to be the best on the basis of comparative analysis of root-mean-square error (RMSE) at zero epoch. A significant correlation between the model-predicted and Soil-Plant Analysis Development (SPAD) meter-measured relative chlorophyll contents was obtained when the mean brightness ratio (rgb) parameters were used. Compared to a red (R), green (G), and blue (B) color space model, the rgb model exhibited better performance with a significant correlation (R ² = 0.85). Incorporation of photometric features, such as luminosity (L), blue (B)/L, and green (G)/L, with rgb failed to improve the performance of the network. The developed Intelligent image analysis (IIA) system was able to estimate in real time the chlorophyll content of in vitro-regenerated leaves for assessment of plant nutrient status during micropropagation.     

Analysis of quantitative trait loci underlying the traits related to chlorophyll content of the flag leaf in rice

A population of 117 doubled haploid (DH) lines derived from the cross of Zhaiyeqing 8 (indica) x Jingxi 17 (japonica) was employed to map quantitative trait loci (QTL) underlying four physiological traits related to chlorophyll contents of the flag leaf. There were significantly positive correlations among chlorophyll a, chlorophyll b and chlorophyll a+ b content. Chlorophyll a/b ratio was significantly negatively correlated with chlorophyll b content. These four traits were normally distributed with transgressive segregation, suggesting that they were controlled by multiple minor genes. A total of 11 QTLs were detected for the four traits and they lay on six chromosomes. Each of them explained 9.2%-19.6% of the phenotypic variations, respectively. Of these, two QTLs controlling chlorophyll a content were mapped on chromosomes 2 and 5; four QTLs underlying chlorophyll b content were mapped on chromosomes 2, 3, 5 and 9; three QTLs underlying chlorophyll a+b amount were mapped on chromosomes 3, 5 and 9; two QTLs under-lying chlorophyll a/b ratio were mapped on chromosomes 6 and 1 1. The intrinsic relationship among the four traits and the practical implication in rice breeding are discussed.     

淹水胁迫下棉花叶片高光谱特征及叶绿素含量估算模型

为即时监测淹水胁迫下棉花功能叶叶绿素含量,确立叶绿素含量与单叶光谱特征参数的定量关系,本文以遮雨棚内蕾期淹水胁迫下的盆栽棉花为研究对象,淹水后每 3 d对棉花叶片进行测定、取样,综合分析叶绿素含量与高光谱特征参数的相关性,并构建和验证叶绿素含量的估算模型.结果表明: 随着淹水胁迫程度的加重,叶片叶绿素含量下降;叶片原始光谱反射率、一阶微分光谱反射率分别在580、697 nm波段附近与叶绿素含量呈显著负相关;利用差值指数和归一化指数建立的估算模型优于单波段线性模型,其中以植被指数(DR₆₉₇-DR₇₃₈)/(DR₆₉₇+DR₇₃₈)为自变量建立的模型棉花单叶叶绿素含量估算值与实测值拟合度最好,拟合系数为0.814,可用于淹水胁迫下棉花单叶叶绿素含量的估测.     

不同叶色红栌叶片中色素含量、酶活性及内含物差异的研究

为明确秋季不同叶色美国红栌叶片的生理差异,以秋季同一植株上红色、中间色、绿色三种颜色的美国红栌叶片为试材,测定了叶片中色素物质含量、酶活性以及叶片可溶性内含物的含量,结果表明：在红色叶片中,叶绿素含量较低,PAL、POD酶活性较高,花青素苷/叶绿素的比值较大,从而使叶色显现红色;而在绿色叶片中,叶绿素含量较高,PAL、POD酶活性较小,花青素苷/叶绿素的比值较低,叶片显现绿色。通过可溶性内含物测定可知,在红色叶片中的可溶性糖和蛋白质含量相对较高,均与花青素苷/叶绿素的比值达到显著相关水平,表明这些内含物的积累有利于花色素苷的合成。     

不同种源黄连木秋季色素含量与叶色参数的关系

该研究以陕西汉中、河南林州、河北涉县和北京中国科学院植物研究所4个种源黄连木(Pistacia chinensis Bunge)的苗木为对象,用分光光度计和色差仪对其叶绿素、类胡萝卜素、花色素苷含量及叶色参数(L*、a*、b*)进行了测定分析,探讨不同种源苗木秋季叶色变化规律及差异,揭示黄连木叶色呈现与叶片色素含量之间的内在关联,为筛选适合城市绿化的优良黄连木种源提供依据。结果表明:(1)在秋季叶片转色期,随着时间的推移,4个种源黄连木叶片的叶绿素、类胡萝卜素和花色素苷含量的比例呈现不同的变化趋势,其中:河北种源的花色素苷含量较高,叶片呈现红色;陕西种源叶绿素含量较高,叶片呈现绿色的时间较长;河南、北京种源处于两者之间。(2)各个种源黄连木的叶色参数a*值(红/绿)均与花色素苷含量呈正相关关系,与叶绿素含量呈负相关关系,且相关系数均达到显著水平(P0.05),各个种源叶色参数L*值(光泽明亮度)也与叶绿素含量间表现出显著或极显著的正相关性。研究发现,河北种源黄连木秋季的叶色最红,陕西种源黄连木叶片呈现绿色的时间最长;色差仪的应用实现了叶色和各色素含量间量化的关系。     

紫花含笑花被呈色过程中色素含量与超微结构的变化特征

为了探究色素含量以及细胞结构在紫花含笑花被呈色过程中的作用机理,该研究以绿色和紫色花被为材料,测定其花被色素含量,运用逐步回归方程分析花被呈色与色素含量的关系,采用石蜡切片及超薄切片技术观察花被细胞超显微结构变化。结果表明:(1)在紫花含笑花被呈色过程中,紫色花被表面明度L^*值降低,a^*值上升,b^*值降低;花被花青素苷的积累量以及类胡萝卜素和类黄酮等含量增加,同时伴随着叶绿素的降解及其含量降低。(2)a^*与花青素、类黄酮、类胡萝卜素等色素含量以及花青素/类黄酮、花青素/叶绿素呈显著正相关关系,b^*与叶绿素含量和花青素/类胡萝卜素呈显著正相关关系。(3)在细胞结构上,随着花被由绿转紫,其上表皮细胞由扁平型向圆锥凸起型变化,单个细胞长宽比增大,细胞垂周壁出现褶皱,紫色花被上表皮结构向增加入射光吸收面积变化;液泡体积增大与叶绿体向有色体转化是主要的细胞器变化。研究发现,花被呈色是多因素作用的结果,花青素含量的产生与积累以及类胡萝卜素和类黄酮等含量增加辅助增色可能是紫花含笑呈紫色的主要...     

西花蓟马繁殖力与寄主营养物质的关系

研究了西花蓟马Frankliniella occidentalis(Pergande)在6种蔬菜寄主上的繁殖力,同时测定了寄主植物叶片的可溶性糖、可溶性蛋白质和叶绿素含量,并分析了它们之间的相关性。研究结果表明,西花蓟马繁殖力与叶片可溶性蛋白质含量存在正相关关系,与叶片的可溶性含糖量、叶绿素含量及可溶性蛋白质含量与可溶性含糖量比例的相关关系不显著。     

Increase of the chlorophyll fluorescence ratio F690/F735 during the autumnal chlorophyll breakdown

Summary The chlorophyll content and the fluorescence induction kinetics at two wavelengths (690 nm and 735 nm) have been measured in leaves of nine common broadleaf tree species during the autumnal chlorophyll breakdown. The ratio of the chlorophyll fluorescence maxima F690/F735 was determined at fluorescence maximum (fm) and at steady-state conditions (fs) by the laser-induced fluorescence emission using the two-wavelength fluorometer. The ratio F690/F735 increases with the leaf discolouring during the autumnal chlorophyll breakdown. The relationship between the chlorophyll content and the ratio F690/F735 can be expressed by a power function (curvilinear relationship) which is valid for all the species examined. In most cases the ratio F690/F735 measured in the upper leaf side is lower than that in the lower leaf side, but the trend is the same along the decreasing chlorophyll content. The ratio F690/F735 is always higher at maximum fluorescence than at steady-state fluorescence in the upper as well as lower leaf side and these values are well fitted in a linear correlation. This study confirms the usefulness of the ratio F690/F735 as a suitable non-destructive indicator of the in-vivo chlorophyll content, especially at medium and low chlorophyll content.     

Analysis of quantitative trait loci underlying the traits related to chlorophyll content of the flag leaf in rice

A population of 117 doubled haploid (DH) lines derived from the cross of Zhaiyeqing 8 (indica)× Jingxi 17 (japonica) was employed to map quantitative trait loci (QTL) underlying four physiological traits related to chlorophyll contents of the flag leaf. There were significantly positive correlations among chlorophyll a, chlorophyll b and chlorophyll a + b content. Chlorophyll a/b ratio was significantly negatively correlated with chlorophyll b content. These four traits were normally distributed with transgressive segregation, suggesting that they were controlled by multiple minor genes. A total of 11 QTLs were detected for the four traits and they lay on six chromosomes. Each of them explained 9.2%–19.6% of the phenotypic variations, respectively. Of these, two QTLs controlling chlorophyll a content were mapped on chromosomes 2 and 5; four QTLs underlying chlorophyll b content were mapped on chromosomes 2, 3, 5 and 9; three QTLs underlying chlorophyll a + b amount were mapped on chromosomes 3, 5 and 9; two QTLs underlying chlorophyll a/b ratio were mapped on chromosomes 6 and 11. The intrinsic relationship among the four traits and the practical implication in rice breeding are discussed. __________ Translated from Journal of Wuhan University (Science Edition), 2006, 52(6): 751–756 [译自: 武汉大学学报(理学版)]     

不同海拔短肋羽藓叶绿素含量与光合效率研究

以云南高黎贡山百花岭不同海拔梯度(1500～3100 m)的短肋羽藓(Thuidium kanedae)为材料,通过测定其叶绿素含量和实际光合量子产量,研究其光合特性随海拔梯度变化的规律。结果表明,短肋羽藓的实际光合量子产量Y(Ⅱ)与叶绿素总量、叶绿素a、叶绿素b含量及叶绿素a/b比值之间都呈负相关关系。随着海拔升高,其叶绿素总含量、叶绿素a及叶绿素b含量都呈先升高而后下降的“单峰”分布模式,但随着海拔进一步升高,叶绿素总含量及叶绿素a含量变化不明显,而叶绿素b含量呈现明显上升趋势。叶绿素a/b比值随着海拔升高先呈现“单峰”分布模式,然后随着海拔继续升高呈现缓慢下降的趋势。实际光合量子产量Y(Ⅱ)整体表现出随海拔升高而增加的趋势,并在高海拔处出现急剧上升的趋势。本研究为揭示苔藓植物在不同海拔梯度下适应生态环境的生理机制提供参考。     

Relationships between carotenoid composition and growth habit in British plant species

The pigment composition of leaves from a number of different plant species collected from field sites in the region of Sheffield, UK, have been compared using high-performance liquid chromatography. Expression of pigment content per unit leaf area was dominated by variation in the total leaf chlorophyll. Neither chlorophyll per unit area nor the chlorophyll a/b ratio were found to be correlated with the habitat from which the plants originated. When the amounts of different carotenoids were expressed relative to the total carotenoid pool, it was found that whilst neither total carotene (α- +β-carotene) nor neoxanthin correlated with ability to grow in shade, the leaf content of both lutein and the total xanthophyll cycle carotenoids (zeaxanthin, anther-axanthin and violaxanthin) did, with lutein content being high in shade species and xanthophyll cycle intermediates low. There was a strong negative correlation between the relative amounts of each of these groups of carotenoids. The ratio of lutein to xanthophyll cycle carotenoids was strongly correlated to an index of shade tolerance.     

Remote-sensing of benthic chlorophyll: should ground-truth data be expressed in units of area or mass?

During low tide, field spectrometric data (350-1050 nm) were acquired from intertidal mudflats in the upper reaches of Sydney Harbour, after which samples of sediment were taken using a small contact core. A total of 103 spectra/sediment pairs of samples were acquired. In the laboratory, amounts of chlorophyll in the contact cores were determined spectrophotometrically. The proportion of fine sediment (<63 μm), a major factor influencing bulk-density of sediment, was measured.The relationship between the remotely sensed observations and the amounts of chlorophyll was investigated by comparing chlorophyll expressed as a mass per unit area (area-normalised values) and as a mass per unit mass (content or weight-normalised values) with: (i) reflectance between 350 and 1050 nm and (ii) a ratio of reflectance at 562 and 647 nm (R562/R647). The reflectance at wavelengths >400 nm were found to be more tightly correlated with area-normalised chlorophyll (maximal correlation=−0.51 at 666 nm) than with weight-normalised chlorophyll (maximal correlation=−0.41 at 664 nm). The relationship between R562/R647 and area-normalised chlorophyll was stronger (R²=0.66) than for weight-normalised chlorophyll (R²=0.47). The residuals from the regression of weight-normalised chlorophyll on R562/R647 were strongly correlated with the mass per unit mass of sediment that was <63 μm. There was no such relationship for residuals from the regression of area-normalised chlorophyll on R562/R647. The results demonstrate conclusively that chlorophyll measured as mass per unit mass of sediment, is inappropriate for ground-truthing remotely sensed observations of chlorophyll in intertidal benthic sediments.     

杂种杨无性系的光系统Ⅱ放氧活性、光合色素及叶绿体超微结构对光胁迫的响应

 用氧电极仪、红外CO2气体分析仪及叶绿素荧光仪,结合透射电镜技术对几个杂种杨无性系在光胁迫下的光系统Ⅱ活性、光合色素及叶绿体超微结构进行了测定。随着预处理光强的增加,各无性系叶片的净光合速率和光系统Ⅱ放氧活性明显下降。二倍体无性系B11的光系统Ⅱ最大光化学效率（Fv/Fm）及实际光化学效率［(Fm′-F)/Fm′］高于两个三倍体无性系B346和B342。强光下三倍体无性系B346的非光化学淬灭远高于B342和二倍体无性系B11。叶绿素含量和光合速率没有明显的线性关系,叶绿素a/b含量在自然光胁迫下存在季节变化,受强光胁迫2个三倍体无性系的叶绿素a/b增大。预处理光强PFD超过3 000 μmol photons·m-2·s-1,各无性系的基粒类囊体片层结构遭到破坏,但二倍体无性系的类囊体片层结构受破坏程度较三倍体无性系轻。叶绿体蛋白合成抑制剂（硫酸链霉素,SM）可加剧叶绿体超微结构的破坏。     

Ven、K型小麦雄性不育系春性F1杂种的优势表现及与叶绿素含量和同工酶谱的关系

利用Ven、K型小麦雄性不育系与北方春性普通小麦杂交,研究了Ven、K型春性F1杂种的优势表现及其与叶绿素含量、同工酶谱的关系。结果表明：Ven型和K型杂种F1存在显著的个体杂种优质,在主要农艺性状中尤其以单株穗数、千粒重、单株生产力的杂种优势明显,其中单株穗数对单株生产力的贡献最大;相关分析表明：单株生产力与单株穗数呈正相关,相关系数为0.7122;与每穗粒数和千粒重的相关系数分别为－0.1183和0.4941。灌浆期旗叶、倒二叶叶绿素含量与千粒重的相关性最大,呈正相关。杂种F1优势强的组合其种子和苗期的过氧化物同工酶和酯酶的酶谱一般呈双亲互补型或“杂种型”酶带,该特征可作为杂种优势预测的指标。     

Fluctuations in pelagic fish density linked to ambient conditions

Animal behaviour has been broadly studied for its social or functional aspects but less often for understanding the interactions between organisms and their ambient conditions. A pilot observational study was performed by means of underwater camera to investigate the correlation between environmental factors and the density of fish in the epipelagic habitat of a European temperate reservoir. Explored factors confirmed a positive correlation between water temperature and all observed categories: nonpredatory, predatory, single nonpredatory and schooling fish, as well as freshwater bream and roach. On the other hand, de-trended solar irradiance was found to be negatively correlated with density of nonpredatory fish, freshwater bream and European perch. Sunshine duration was negatively correlated with the density of predatory fish. Precipitation a showed positive relationship with single nonpredatory fish and European perch, whereas wind strength had a negative relationship with density of schooling fish. Furthermore, density of predatory fish was positively correlated with density of single nonpredatory fish and counts of observed clusters. Altogether, findings indicate that fish density is correlated with abiotic factors and the occurrence of predators. This suggests that more ecologically complex studies should be encouraged for better understanding of ecological interactions that drive the structure of aquatic ecosystems.     

苦荞重组自交系群体籽粒黄酮含量与产量性状分析

为揭示影响苦荞籽粒黄酮含量的主要因素,获得高黄酮含量的苦荞种质,该文以"小米荞×晋荞2号"重组自交系(RILs)群体为材料,采用酶标仪检测籽粒黄酮含量,调查了百粒重、籽粒长宽比、株高、分枝数及籽粒产量等性状的变异,并探究籽粒黄酮含量与产量性状之间的相关关系,比较了不同粒色、不同粒型苦荞株系的籽粒黄酮含量的差异,基于聚类...     

Iron:phosphorus ratio in surface sediment as an indicator of phosphate release from aerobic sediments in shallow lakes

Analysis of Danish lakes showed that both mean winter and mean summer concentrations of lake water total phosphorus in the trophogenic zone correlated negatively with the total iron to total phosphorus ratio (Fe:P) in surface sediments. No correlation was found between the water total phosphorus concentration and either the sediment phosphorus concentration alone or with sediment calcium concentration. The increase in total phosphorus from winter to summer, which is partly a function of net internal P-loading, was lowest in lakes with high Fe:P ratios in the surface sediment.A study of aerobic sediments from fifteen lakes, selected as representative of Danish lakes with respect to the sediment Fe and phosphorus content, showed that the release of soluble reactive phosphorus was negatively correlated with the surface sediment Fe:P ratio. Analysis of phosphate adsorption properties of surface sediment from 12 lakes revealed that the capability of aerobic sediments to buffer phosphate concentration correlated with the Fe:P ratio while the maximum adsorption capacity correlated with total iron. Thus, the Fe:P ratio may provide a measure of free sorption sites for orthophosphate ions on iron hydroxyoxide surfaces.The results indicate that provided the Fe:P ratio is above 15 (by weight) it may be possible to control internal P-loading by keeping the surface sediment oxidized. Since the Fe:P ratio is easy to measure, it may be a useful tool in the management of shallow lakes.