叶绿素荧光分析技术及应用进展

叶绿素荧光动力学技术被称为研究植物光合功能的快速、无损伤探针,已逐渐在环境胁迫对植物光合作用影响研究方面得到应用,随着叶绿素荧光分析技术的进一步发展,其应用领域和研究空间将进一步拓展.本文介绍了叶绿素荧光分析的基本原理,综述了叶绿素荧光分析技术的应用研究进展.     

封面故事

细胞自噬是广泛存在于真核细胞中的生命现象,是生物在其发育、老化过程中都存在的一个净化自身多余或受损细胞结构的共同机制。在植物发育、逆境胁迫、饥饿及衰老的过程中,细胞自噬具有确定的作用。叶绿素荧光分析技术是近几年来发展起来的一种探测植物光合作用生理状况的新技术。荧光与植物的光合作用能力、受胁迫状况、生     

盐胁迫对植物叶绿素荧光影响的研究进展

盐胁迫是制约植物生长发育的主要非生物胁迫之一, 研究植物的耐盐机理对开发和有效利用盐碱地有重要的意义。叶绿素荧光动力技术作为研究植物光合生理状况及植物与逆境胁迫关系的理想方法, 可表明外界胁迫环境对植物光合器官的伤害程度。通过总结性阐述盐胁迫对植物叶绿素荧光的影响, 分别从盐分类型、植物类型、光照强度以及盐旱交互作用等方面分析了植物叶绿素荧光对盐胁迫的响应, 进而反映盐胁迫对植物光合能力的影响程度, 并提出增强植物抗盐性的途径, 包括施加外源物质、利用转基因技术、真菌的协同效应和培育耐盐品种。最后对叶绿素荧光动力技术在抗盐胁迫的运用前景进行了展望, 提出了当前研究需要解决的问题, 旨在为提高植物耐盐能力提供一定的理论依据。     

地形和土壤特性对亚热带常绿阔叶林内植物功能性状的影响

植物功能性状与环境之间的关系是功能性状研究的核心问题.为了探讨地形和土壤特性的差异对亚热带常绿阔叶林植物功能性状的影响,并找到影响古田山亚热带常绿阔叶林植物形态和生理性状的主要环境驱动因子,我们于2008年和2009年夏天在古田山国家级自然保护区内24 ha大型监测样地测定了147个样方中115种常见木本植物的功能性状.所测性状包括3个生理生态性状(叶绿素含量、叶绿素荧光参数Fv/Fm和PIABS和枝条比导率)和4个形态性状(气孔密度、叶厚度、比叶面积和叶长/宽比).结合地形数据(平均海拔、凹凸度、坡度和坡向)和土壤数据(含水量、全氮含量、全磷含量、全碳含量和pH值),分析影响这些功能性状的主要驱动因子.排序结果显示,叶绿素含量随海拔和凹凸度的上升而下降,但与土壤中的氮含量和水分含量旱现正相关关系.由于古田山土壤呈酸性,土壤磷素缺乏,叶绿素荧光参数Fv/Fm和PIABS与土壤氮、磷含量呈现显著的负相关.枝条比导率与土壤含水量具有较为显著的正相关关系.比叶面积与海拔呈现正相关关系.研究结果表明,在小尺度上,海拔和凹凸度是影响亚热带常绿阔叶林植物功能性状晟关键的两个地形因子,而土壤含水量和伞氮含量是影响该地植物功能性状最主要的土壤因子.然而,由于土壤中磷的缺乏,诸如植物光合作用等一些重要的生理过程受到影响,使得某些性状与环境因子之间呈现出不同寻常的相关关系.     

叶绿素荧光主动与被动联合观测应用前景

叶绿素荧光是研究植物光合生理机制、量化植被光合作用时空格局以及准确理解气候变化背景下陆地生态系统生产力的关键。然而, 目前对于叶绿素荧光主动与被动联合观测的研究还较少。该文对比了叶绿素荧光主动观测与被动观测的优缺点, 展示了叶片尺度和冠层尺度主动与被动联合观测的仪器设备组成, 探讨了主动与被动联合观测在探索叶绿体尺度-叶片尺度-冠层尺度能量在光合、荧光以及热耗散中的分配, 阐明叶绿素荧光与总初级生产力的关联机制, 验证星基日光诱导叶绿素荧光, 解译叶绿素荧光光谱形状4个方面的应用前景。综上, 叶绿素荧光的主动与被动联合观测对于揭示各尺度上荧光与光合作用之间的关联机制, 改善全球尺度植被生产力模型至关重要。     

用叶绿素荧光分析技术鉴定植物抗寒性的剖析

在分析叶绿素荧光技术原理的基础上,从低温条件下叶绿素荧光参数的变化与处理温度、季节的关系、叶绿素荧光参数的变化与其它抗寒指标的相关性以及可用于筛选与鉴定抗寒植物的荧光参数3个方面介绍这一技术在抗寒植物筛选与鉴定中的应用现状,并指出这一技术的可取之处和存在问题。     

植物反射光谱对水分生理变化响应的研究进展

实时、无损伤地探测植物的水分及生理变化是高光谱遥感的深层次应用。由水分胁迫引发的植物一系列反射光谱响应体现了碳-氮-水耦合作用的结果。以往的研究大多集中于单一因素的响应, 而忽略了多因素交互作用。该文综述和分析了植物水分状况变化引起的直接和间接光谱响应机制, 包括植物水分含量、色素、养分状况、光合作用和叶绿素荧光指标的光谱响应及其内在的关联, 探讨了反射光谱在探测植物水分生理活动应用中的主要方法与最新技术, 并指出碳-氮-水多指标、多时空尺度的综合分析对于估测植被生产力及其对气候变化的响应具有重要意义。     

植物叶色白化研究进展

植物叶色白化是一类明显和常见的叶绿素缺失突变,在植物光合作用机理、激素生理、核一质基因组相互作用、遗传育种等理论研究和实际应用方面均具有无可替代的价值。主要综述了近年来国内外有关白化突变体与叶绿素合成的关系、影响白化的内外因素、相关基因的克隆以及克服白化的途径等方面的研究进展,旨在为深入研究植物叶色白化突变提供参考。     

快速叶绿素荧光动力学及其在植物抗逆生理研究中的应用

快速叶绿素荧光动力学是研究光合原初反应的无损探针,可监测光合反应的多个事件,并能反应植物的生理状况,可以应用于环境物理及化学条件胁迫对植物的影响(如化肥及除草剂的使用;CO2、O2、O3浓度的变化及温度、光强度胁迫等);也应用于农业生产(如确定耕作方式及日常管理;除草剂、杀虫剂及激素的使用;品种选优等);更是光合生理研究的便捷工具。介绍了快速叶绿素荧光动力学中的JIP-test分析方法,并从光、水分、温度、盐等多个方面介绍了JIP-test方法在植物抗逆生理研究中的应用。     

植物叶片荧光光谱测绘技术探讨

植物叶绿素荧光是研究植物生理变化、环境胁迫效应和光合作用原初过程的一个无损探针。愈来愈多的作者已认识到直接研究植物叶片荧光的意义。但是,由于植物荧光弱、叶表面光散射强以及荧光重吸收等多方面因素,要得到一幅准确的荧光光谱是极为困难的。而另一方面,测绘到的荧光光谱是否能最大程度地反映叶片荧光发射情况,会直接影响研究结论。这样光谱测     

柚木无性系光合生理特征与生长综合评价

为了掌握柚木无性系光合生理与生长性状遗传变异规律,筛选出优良无性系。基于柚木无性系测定林,测定选自不同种源的19个无性系+1个对照的气体交换、叶绿素荧光参数和生长性状,分析柚木无性系光合生理特征及其与生长的关系。结果表明：气体交换、叶绿素荧光参数和树高在柚木不同无性系间、不同种源间都存在极显著差异,除了PSⅡ最大光化学效率（F_v/F_m）,其他各性状都具有丰富的遗传变异,但净光合速率等光合生理参数与树高生长没有显著的相关性。结论：早期选出71-7、FS3为光合速率高和树高生长好的优良无性系,起源于印度的柚木资源可作为今后高光效育种选择的重点,需要对光合生理和生长测定进行综合评价才能准确地筛选优良无性系。     

Chlorophyll fluorescence of the trunk rather than leaves indicates visual vitality in Eucalyptus saligna

Leaf chlorophyll fluorescence has been used to assess physiological stress effects in trees for at least 30?years. This paper describes a novel method for indicating tree vitality in Eucalyptus saligna Sm using bark chlorophyll fluorescence. A visual vitality index was successfully verified by comparing it with growth measurements such as total leaf area and above ground biomass. Bark and leaf chlorophyll fluorescence were then compared with the visual vitality index in spring, summer and autumn. The relationship between leaf chlorophyll fluorescence and the visual vitality index was weak, even with the one parameter that did show a relationship. There was good evidence for a statistical relationship between bark chlorophyll fluorescence and visual vitality, which may become a useful tool for tree vitality assessments in this species and possibly other tree species.     

森林次生演替优势种苗木的光可塑性比较研究

本文从研究苗木的叶绿素含量和ＲｕＢＰ羧化酶活性随着光环境的变化而发生改变的规律出发,来探讨森林次生演替优势种苗木的光可塑性大小和对弱光环境的适应能力。各种苗木的叶绿素含量都随着光强度变弱而增加,但如果较长时间生长在弱光环境中,由于叶绿素的合成小于分解,其含量也会逐渐变小。不同的演替阶段优势种苗木的叶绿素含量的增加或减少在量上有一定的区别。以叶绿素含量随着光环境变化的测定值为指标,用模糊数学分析的结果表明,苗木的耐荫性大小顺序是演替后期种（黄果厚壳桂Ｃｒｙｐｔｏｃａｒｙａｃｏｎｃｉｎｎａ）＞演替过渡种（藜蒴Ｃａｓｔａｎｏｐｓｉｓｆｉｓａ）＞演替过渡种（荷木Ｓｃｈｉｍａｓｕｐｅｒｂａ）＞演替先锋树种（马尾松Ｐｉｎｕｓｍａｓｏｎｉａｎａ）;且藜蒴和荷木很接近,稍靠近黄果厚壳桂。马尾松和荷木的ＲｕＢＰ羧化酶活性随着生长环境的光强度的增加,其活性有所增加;但黄果厚壳桂的相应值是在每日直照光１ｈ的光环境中最高。除马尾松外,演替过渡种和后期种的苗木都是在每日直照光１ｈ的光环境中生长最好,这和每日短期照光提高ＲｕＢＰ羧化酶活性的（与没有直照光的环境相比较）同时又不分解叶绿素、不降低其含量有密切的关系。     

森林次生演替优势种苗木的光可塑性比较研究

本文从研究苗木的叶绿素含量和RuBP羧化酶活性随着光环境的变化而发生改变的规律出发,来探讨森林次生演替优势种苗木的光可塑性大小和对弱光环境的适应能力。各种苗木的叶绿素含量都随着光强度变弱而增加,但如果较长时间生长在弱光环境中,由于叶绿素的合成小于分解,其含量也会逐渐变小。不同的演替阶段优势种苗木的叶绿素含量的增加或减少在量上有一定的区别。以叶绿素含量随着光环境变化的测定值为指标,用模糊数学分析的结果表明,苗木的耐荫性大小顺序是演替后期种(黄果厚壳桂Cryptocarya concinna)>演替过渡种(藜蒴 Castanopsis fissa)>演替过渡种(荷木 Schima superba)>演替先锋树种(马尾松Pinus massoniana);且藜蒴和荷木很接近,稍靠近黄果厚壳桂。马尾松和荷木的RuBP羧化酶活性随着生长环境的光强度的增加,其活性有所增加;但黄果厚壳桂的相应值是在每日直照光1h的光环境中最高。除马尾松外,演替过渡种和后期种的苗木都是在每日直照光1 h的光环境中生长最好,这和每日短期照光提高RuBP羧化酶活性的(与没有直照光的环境相比较)同时又不分解叶绿素、不降低其含量有密切的关系。     

华山松叶绿素荧光诱导动力学参数的地理变异及其与树高生长的关系

 利用叶片体内荧光测定技术,检测了南北5个地区华山松种源的荧光诱导动力学参数。结果表明,南方种源具有较强的最大荧光(Fm)和可变荧光(Fv),其光系统Ⅱ(PSⅡ)的潜在活性(Fv／Fo)与原初光能转换效率(Fv／Fm)也明显高于北方种源,并与其树高生长成正相关。另一方面,北方种源的非光化学荧光猝灭系数(QN)比南方种源要大,与它们的树高生长成反相关,而与纬度成正相关。这些实验结果说明,利用荧光诱导动力学技术,可以检测华山松的生长量潜力,在华山松种源选择中有较大的应用前景。     

Rapid recovery of tropical forest diversity and structure after shifting cultivation in the Philippines uplands

Shifting cultivation is a widespread land‐use in the tropics that is considered a major threat to rainforest diversity and structure. In the Philippines, a country with rich biodiversity and high rates of species endemism, shifting cultivation, locally termed as kaingin, is a major land‐use and has been for centuries. Despite the potential impact of shifting cultivation on forests and its importance to many people, it is not clear how biodiversity and forest structure recover after kaingin abandonment in the country, and how well these post‐kaingin secondary forests can complement the old‐growth forests. We investigated parameters of forest diversity and structure along a fallow age gradient in secondary forests regenerating after kaingin abandonment in Leyte Island, the Philippines (elevation range: 445–650 m asl). We first measured the tree diversity and forest structure indices in regenerating secondary forests and old‐growth forest. We then measured the recovery of tree diversity and forest structure parameters in relation to the old‐growth forest. Finally, using linear mixed effect models (LMM), we assessed the effect of different environmental variables on the recovery of forest diversity and structure. We found significantly higher species density in the oldest fallow sites, while Shannon’s index, species evenness, stem number, basal area, and leaf area index were higher in the old‐growth forest. A homogeneous species composition was found across the sites of older fallow age. Multivariate analysis revealed patch size as a strong predictor of tree diversity and forest structure recovery after shifting cultivation. Our study suggests that, secondary forests regenerating after shifting cultivation abandonment can recover rapidly. Although recovery of forest structure was not as rapid as the tree diversity, our older fallow sites contained a similar number of species as the old‐growth forest. Many of these species are also endemic to the Philippines. Novel and emerging ecosystems like tropical secondary forests are of high conservation importance and can act as a refuge for dwindling tropical forest biodiversity.     

Photosynthetic Responses to Light for Rainforest Seedlings Planted in Abandoned Pasture, Costa Rica

Efforts to reforest tropical pasture with native tree species have increased in recent years, yet little is known about the physiology of most tropical trees. The goal of this study was to assess the effect of habitat on photosynthetic responses to light for seedlings of four native rainforest species (Calophyllum brasiliense, Ocotea glaucosericea, Ocotea whitei, and Sideroxylon portoricense) planted to facilitate tropical rainforest recovery in southern Costa Rica. Seedlings were planted in primary forest, in open abandoned pasture, and in the shade of remnant trees within the pasture. Growth, morphology, photosynthetic gas exchange responses to light, and chlorophyll fluorescence (an indication of the integrity of photosynthetic processes) were measured in the three habitats. Height and leaf area were generally greater for seedlings in tree shade compared to those in the forest and open pasture. Photosynthetic rates were higher for plants in open pasture and tree shade compared to those in the forest for two of the four species. Chlorophyll fluorescence results indicated flexibility in the photosynthetic processing of light energy that may help plants tolerate the bright light of the pasture. This study demonstrates that, for certain species, seedlings under remnant pasture trees do not exhibit the level of photosynthetic stress experienced in open abandoned pasture. Seedling responses to light, in combination with other factors such as increased nutrient input through litterfall, help explain the enhanced growth of seedlings under remnant pasture trees. Planting seedlings under remnant trees may increase the success of future efforts to restore tropical forest in abandoned agricultural land.     

Remote sensing of heterogeneity in photosynthetic efficiency, electron transport and dissipation of excess light in Populus deltoides stands under ambient and elevated CO2 concentrations, and in a tropical forest canopy, using a new laser-induced fluorescence transient device

Determining the spatial and temporal diversity of photosynthetic processes in forest canopies presents a challenge to the evaluation of biological feedbacks needed for improvement of carbon and climate models. Limited access with portable instrumentation, especially in the outer canopy, makes remote sensing of these processes a priority in experimental ecosystem and climate change research. Here, we describe the application of a new, active, chlorophyll fluorescence measurement system for remote sensing of light use efficiency, based on analysis of laser‐induced fluorescence transients (LIFT). We used mature stands of Populus grown at ambient (380 ppm) and elevated CO₂ (1220 ppm) in the enclosed agriforests of the Biosphere 2 Laboratory (B2L) to compare parameters of photosynthetic efficiency, photosynthetic electron transport, and dissipation of excess light measured by LIFT and by standard on‐the‐leaf saturating flash methods using a commercially available pulse‐modulated chlorophyll fluorescence instrument (Mini‐PAM). We also used LIFT to observe the diel courses of these parameters in leaves of two tropical forest dominants, Inga and Pterocarpus, growing in the enclosed model tropical forest of B2L. Midcanopy leaves of both trees showed the expected relationships among chlorophyll fluorescence‐derived photosynthetic parameters in response to sun exposure, but, unusually, both displayed an afternoon increase in nonphotochemical quenching in the shade, which was ascribed to reversible inhibition of photosynthesis at high leaf temperatures in the enclosed canopy. Inga generally showed higher rates of photosynthetic electron transport, but greater afternoon reduction in photosynthetic efficiency. The potential for estimation of the contribution of outer canopy photosynthesis to forest CO₂ assimilation, and assessment of its response to environmental stress using remote sensing devices such as LIFT, is briefly discussed.     

Application of chlorophyll fluorescence in ecophysiology

Summary In vivo chlorophyll fluorescence measurements have become a valuable tool in ecophysiology. Fluorescence emission spectra are influenced by the reabsorption of the tissue and indicate the composition of the antenna system and are influenced by the chlorophyll content per leaf area. The fluorescence induction kinetics (Kautsky effect) can be used to study photosynthetic activity. These rapid, non-destructive methods can be applied for ecophysiological field research to check the vitality of plants and to document stress effects on the photosynthetic apparatus. The Rfd-values (Rfd=fd/fs), the ratio of the fluorescence decrease (fd) to the steady state fluorescence (fs), can be taken as a rapid vitality index of the leaves and trees. We here describe fundamental chlorophyll fluorescence results of leaves which are needed for the interpretation of in vivo fluorescence signatures in stress physiology and in the forest dieback research.