Ozone foliar symptoms in woody plant species assessed with ultrastructural and fluorescence analysis

This paper compares the responses to ozone in five woody species: Fagus sylvatica (FS), Acer pseudoplatanus (AP), Fraxinus excelsior (FE), Viburnum lantana (VL) and Ailanthus altissima (AA). The hypothesis being tested was that the strategies that plants adopt to resist oxidative pressure are species-specific. The study was carried out on field grown plants in an area in Northern Italy characterized by elevated levels of ozone pollution. The observations were made both at ultrastructural (using light and electronic microscopy) and physiological (using chlorophyll a transient fluorescence and microspectral fluorometry) level. Common responses were: the hypersensitive response (i.e. the death of palisade mesophyll cells) and the formation of callose layers separating injured from healthy cells. FS and AP were capable of thickening the palisade mesophyll cell walls. This thickening process involved changes in cell wall chemical structure, evidenced by the accumulation of yellow autofluorescence compounds. Species-specific behaviours were observed with the fluorescence analysis, with special reference to the photochemical de-excitation constant (Kp). This value increased in FE and AP, and decreased in AA. The observed responses are interpreted as adaptative strategies against the ozone stress. The increase of Kp indicates that the reaction centres were working as more effective quenchers.     

Interactions and competition processes among tree species in young experimental mixed forests,assessed with chlorophyll fluorescence and leaf morphology

Chlorophyll a fluorescence (ChlF) and leaf morphology were assessed in two sites in Europe (Kaltenborn, Germany, and Satakunta, Finland) within a forest diversity experiment. Trees at Satakunta, planted in 1999, form a stratified canopy, while in Kaltenborn the trees are 7 years old, with no apparent canopy connection among broadleaf species. The following ChlF parameters from measured OJIP transient curves were examined: F_V/F_M (a proxy for maximum quantum yield); Ψ_Eo (a proxy for efficiency in transferring an electron from reduced QA to the electron transport chain); I‐P phase (a proxy for efficiency of reducing final acceptors beyond PSI); and PI_tot (total performance index for potential energy conservation from photons absorbed by PSII to reduction of PSI end acceptors). At Satakunta F_V/F_M and Ψ_Eo in Betula pendula were higher in monocultures and lower in mixed plots, perhaps due to increasing light availability in mixed plots, which can induce photoinhibition. The opposite trend was observed in Picea abies, which was shaded in mixed plots. At Kaltenborn F_V/F_M decreased in Fagus sylvatica and P. abies in mixed plots due to competition both above‐ and belowground. At Satakunta LMA increased in B. pendula leaves with increasing species richness. Leaf area of ten leaves was reduced in F. sylvatica in mixed plots at Kaltenborn. By up‐scaling the overall fluorescence response to plot level (PI_{tot_plot}), a significant positive correlation with tree diversity was found at Kaltenborn, but not at Satakunta. This could suggest that competition/facilitation processes in mixed stands play a significant role in the early stages of forest establishment, but then tend to be compensated in more mature stands.     

两种生境条件下6种牧草叶绿素含量及荧光参数的比较

昆仑山前山牧场海拔较高, 策勒绿洲海拔相对较低, 两者生境差异较大。以昆仑山前山牧场和策勒绿洲边缘两种不同生境条件下生长的6种牧草: 冰草(Agropyron cristatum)、无芒雀麦(Bromus inermis)、矮生高羊茅(Festuca elata)、披碱草(Elymus dahuricus )、红豆草(Onobrychis pulchella)及和田大叶(Medicago sativa var. luxurians)为试验材料, 研究了不同生境条件下牧草叶片叶绿素含量及叶绿素荧光动力学参数的变化情况。结果显示: (1)在两种生境条件下, 昆仑山前山牧场生境生长的牧草叶绿素a、叶绿素b、总叶绿素的含量明显较高, 生长在策勒绿洲生境的牧草品种叶绿素a/b值较高; (2)昆仑山前山牧场生境牧草最大荧光、光系统II (PSII)最大光化学效率、PSII潜在活性和单位面积反应中心的数量的值明显高于策勒绿洲生境品种, 而初始荧光、单位反应中心吸收的光能、单位反应中心捕获的能量、单位反应中心耗散的能量、荧光诱导曲线初始斜率值则低于策勒绿洲生境品种。因此, 两种生境下环境因子发生了改变, 对牧草产生综合的胁迫作用; 策勒绿洲生境明显对牧草生长产生了抑制, 策勒绿洲生境牧草的色素含量降低以及PSII的机构遭到损坏, 导致反应中心一部分失活或裂解, 剩余有活性的反应中心的效率增加, 昆仑山生境则相对比较适宜牧草生长; 两种生境不同牧草叶绿素含量和叶绿素荧光参数的变化幅度不同。     

8种阔叶树种叶片气体交换特征和叶绿素荧光特性比较

在自然条件下,测定了8种阔叶树种叶片的气体交换参数和叶绿素荧光参数并对其进行比较.结果表明,8种阔叶树种紫玉兰、广玉兰、玉兰、美人梅、铁杆梅、腊梅、红碧桃和紫薇的叶片净光合速率（Pn）、蒸腾速率（Tr）、气孔导度（gs）、瞬时水分利用效率（WUE）和潜在水分利用效率（WUEi）的种间差异达极显著水平（p〈0.01）,指示了不同树种间的光合能力及水分利用能力差别较大.8种阔叶树种叶片的初始荧光（Fo）、可变荧光（Fv）、最大荧光（Fm）和PSⅡ电子传递量子效率（ФPSⅡ）的种间差异极为显著（p〈0.01）,PSⅡ最大光能转换效率（Fv/Fm）、可变荧光与初始荧光之比（Fv/Fo）和非光化学猝灭系数（NPQ）的种间差异也达显著水平（p〈0.05）,说明各树种叶片的PSⅡ原初光能转换效率和潜在活性、PSⅡ电子传递量子效率以及PSⅡ的潜在热耗散能力差别较大,而实际光下最大荧光（F′m）和PSⅡ光能捕获效率（F′v/F′m）的种间差异不显著.3种木兰科植物的Pn、Tr、WUE和WUEi平均值均高于3种蔷薇科植物,说明木兰科植物的光合能力较强,对吸收的光能和水分的利用较高.蔷薇科植物的Fv/Fm、Fv/Fo、ФPSⅡ、F′v/F′m和光化学猝灭系数（qp）平均值均高于木兰科植物,而木兰科植物NPQ较高,表明其PSⅡ的潜在热耗散能力较强,可有效地避免过剩光能对光合机构的损伤.研究还表明3种木兰科植物和3种蔷薇科植物之间的叶绿素荧光参数差异不大,说明同一科属植物叶片的光合能力较为相近.相关分析表明,8种阔叶树种叶片的Pn与Tr、Tr与gs、Fv/Fm与Fv/Fo、ФPSⅡ与F′v/F′m、qp与NPQ均呈极显著正相关（p〈0.01）,Pn与gs呈显著正相关（p〈0.05）,而Tr、gs与WUE、WUEi,Pn与ФPSⅡ,ФPSⅡ与NPQ,F′v/F′m 与 qp、NPQ均呈极显著负相关（p〈0.01）.     

Photosynthetic acclimation to light in woody and herbaceous species: a comparison of leaf structure, pigment content and chlorophyll fluorescence characteristics measured in the field

Acclimation of foliage photosynthetic properties occurs with varying time kinetics, but structural, chemical and physiological factors controlling the kinetics of acclimation are poorly understood, especially in field environments. We measured chlorophyll fluorescence characteristics, leaf total carotenoid (Car), chlorophyll (Chl) and nitrogen (N) content and leaf dry mass per area (LMA) along vertical light gradients in natural canopies of the herb species, Inula salicina and Centaurea jacea, and tree species, Populus tremula and Tilia cordata, in the middle of the growing season. Presence of stress was assessed on the basis of night measurements of chlorophyll fluorescence. Our aim was to compare the light acclimation of leaf traits, which respond to light availability at long (LMA and N), medium (Chl a/b ratio, Car/Chl ratio) and short time scales (fluorescence characteristics). We found that light acclimation of nitrogen content per unit leaf area (N(area)), chlorophyll content per unit dry mass (Chl(mass)) and Chl/N ratio were related to modifications in LMA. The maximum PSII quantum yield (F(v) /F(m)) increased with increasing growth irradiance in I. salicina and P. tremula but decreased in T. cordata. Leaf growth irradiance, N content and plant species explained the majority of variability in chlorophyll fluorescence characteristics, up to 90% for steady-state fluorescence yield, while the contribution of leaf total carotenoid content was generally not significant. Chlorophyll fluorescence characteristics did not differ strongly between growth forms, but differed among species within a given growth form. These data highlight that foliage acclimation to light is driven by interactions between traits with varying time kinetics.     

基于叶绿素荧光参数的小麦叶片叶绿素相对含量估算方法

叶绿素含量是植物学和农业相关研究领域常用的生理指标.叶绿素含量和叶片光合功能密切相关,但是现有的叶绿素含量的测定方法无法实现叶绿素含量和光合功能的同步测定和关联分析.为解决该问题,本研究通过测定35个小麦品种旗叶的SPAD值和叶绿素荧光诱导动力学曲线,分别使用不同时间的快速叶绿素荧光动力学曲线的荧光值,以及33个常用荧...     

Changes in yield ofin-vivo fluorescence of chlorophyll a as a tool for selective herbicide monitoring

Triazines and derivatives of phenylurea, which are often found in outdoor water samples, induce specific changes in the yield of thein-vivo chlorophyll -fluorescence of PSII. These changes are correlated quantitatively with the concentration of the herbicides and can therefore be used to set-up a low-price monitor system. In order to detect selectively the herbicide-sensitive part of the fluorescence emission a pulse amplitude modulated fluorimeter was used. The bioassay system was optimised with respect to test organism, growing and measuring conditions. The relationship between fluorescence yield and herbicide concentrations were experimentally determined for the triazines atrazine and simazine and the phenylurea herbicide DCMU and mathematically fitted (r=0.99). The I₅₀-values were 0.9 µM for DCMU, 2.2 µM for simazine and 3.3 µM for atrazine. The detection limit of about 0.5 µM clearly shows that the sensitivity of this bioassay system is too low to reach the requirements of the drinking water regulation. However, due to its insensitivity against complex water matrices, there is good hope to combine this fluorometric bioassay with a potent herbicide preconcentration method like a solid-phase extraction procedure.Author for correspondence     

Low-temperature limitations of photosynthesis in three tropical Vigna species: A chlorophyll fluorescence study

The temperature-dependence of photosynthesis and chlorophyll (Chl) fluorescence quenching components was studied between 0 and 45°C in three tropical, chilling-sensitive Vigna species and in chilling-tolerant pea. Photosynthesis of the Vigna spp. was approx. 20% more reduced by temperatures between 7 and 30°C than in pea. The latter revealed significant changes in Chl fluorescence parameters at much lower temperature than the Vigna spp. Below 15°C, the reduction state of Q_A increased quickly in pea, while in Vigna already below 30°C, an increase of reduced Q_A was obtained. The analysis of different components of non-photochemical Chl fluorescence quenching (q_N) revealed, that in pea photoinhibitory quenching (q_I) occurred below 13°C. Below ca. 7°C, a sudden breakdown of both q_P and the fast relaxing component of q_N was observed in pea.In Vigna, susceptibility of LHC II phosphorylation or limitation of electron flow by damage to PS I, the PS II reaction centre or the water-splitting system were not responsible for the chilling-sensitivity of photosynthesis between 5 and 30°C. Instead, photosynthesis was gradually limited by an inefficient use of reduction equivalents. This, in turn may increase susceptibilty to photoinhibition, which occurred below 20°C in Vigna. The combined study of q_P and of the different components of q_N allowed the demonstration of the subsequent occurrence of different limiting processes with decreasing temperature in the chilling-sensitive Vigna species.     

快速叶绿素荧光诱导动力学分析在光合作用研究中的应用

JIP-测定(JIP-test)是以生物膜能量流动为基础建立的分析方法.利用该方法可以获得有关光系统Ⅱ的大量信息.文章介绍了快速叶绿素荧光诱导动力学曲线的定义、数据分析方法及相关参数的意义,并举例说明如何利用该方法分析不同环境条件对光合机构主要是PSⅡ的供体侧、受体侧及PSⅡ反应中心的影响.     

Growth control by phytochrome of de-etiolated bean hypocotyls mediated by chlorophyll fluorescence

The elongation of hypocotyls excised from de-etiolated seedlings of beans (Phaseolus vulgaris L. cv. British Wax) is inhibited by light, blue and red irradiations being equally effective. Conditions which decrease chlorophyll fluorescence, such as CO₂-free air, abolish the inhibitory effect of blue irradiation and enhance the inhibition by red light. Conversely, conditions which increase chlorophyll fluorescence, such as a N₂ atmosphere or irradiation through a chlorophyll filter, abolish the inhibitory effect of red light and enhance the inhibition by blue irradiation. The inhibitory effect of blue light is reversible by red irradiation under increased fluorescence as well as by far red. We propose that the chlorophyll fluorescence excited by blue and red irradiations in λ_F > 660 nm and λ_F > 720 nm, respectively, is responsible for the inhibitory effect of blue light and the reduction of the inhibitory effect of non fluorescing red light. Both red and blue wavelengths seem, therefore, to control hypocotyl elongation through phytochrome.     

Photoinhibition in outdoor Spirulina platensis cultures assessed by polyphasic chlorophyll fluorescence transients

Photoinhibition in outdoor cultures of Spirulina platensis was studied by measuring the polyphasic rise of chlorophyll fluorescence transients, which provide information on the primary photochemistry of PSII. The maximum efficiency of PSII photochemustry (F_v/F_m) declined in response to daily increasing irradiance and recovered as daily irradiance decreased. The greatest inhibition (15%) in F_v/F_m was observed at 12:00 hr which responded to the highest irradiance. The absorption flux, the trapping flux, and the electron transport flux per PSII reaction center increased in response to daily increasing irradiance and decreased as irradiance decreased. The daily change in the concentration of PSII reaction centers followed the same pattern as F_v/F_m. However, no significant changes in the probability of electron transport beyond Q_A (Ψ_o) were observed during the day. The results suggest that the decrease in F_v/F_m induced by photoinhibition in outdoor Spirulina cultures was a result of the inactivation of PSII reaction centers. The results also suggest that the measurement of polyphasic fluorescence transients is a powerful tool to study the mechanism of photoinhibition in outdoor Spirulina cultures and to screen strains for photoinhibition tolerance. This revised version was published online in June 2006 with corrections to the Cover Date.     

不同类型喀斯特植物的荧光特征及抗旱性比较

利用叶绿素荧光技术观测了五个不同类型的喀斯特植物翅荚香槐、大盔凤仙、红背叶、牛耳朵和青冈栎在不同作用光强下的光能利用特征,并且对这五种类型植物在PEG诱导水分胁迫下的抗旱性作了比较。结果表明,随着作用光强的增加,这五种植物的光化学反应能力(qP)逐渐降低,非光化学耗散作用(NPQ)明显增加,同时PSⅡ有效光化学效率(Fv′/Fm′)随之下降,导致PSⅡ电子传递量子效率(ΦPSⅡ)也明显下降,但在相同作用光强下这五个类型植物的荧光特征无明显差异。在PEG诱导水分胁迫的实验中,牛耳朵、青冈栎在PEG处理后Fv/Fm变化不明显,而红背叶Fv/Fm值下降程度最大,其次为翅荚香槐、大盔凤仙。由此推测,牛耳朵和青冈栎的耐干旱能力最强,红背叶抗旱能力最弱,翅荚香槐、大盔凤仙居中。     

干旱和复水对入侵植物三裂叶蟛蜞菊叶片叶绿素荧光特性的影响

选取入侵植物三裂叶蟛蜞菊(Wedelia trilobata)及其本地近缘种蟛蜞菊(W. chinensis)为实验材料,比较干旱和复水后二者叶片的叶绿素荧光特性和抗氧化酶活性等生理指标的变化规律,探讨入侵种三裂叶蟛蜞菊对干旱的响应和生态适应性.结果发现,在自然干旱处理过程中,入侵种三裂叶蟛蜞菊与本地种蟛蜞菊相比土壤含水量下降较快,对它们叶片气孔形态的比较发现,干旱胁迫11d后三裂叶蟛蜞菊叶片气孔开度明显大于蟛蜞菊,这可能是导致其失水较快的原因之一.干旱胁迫11d后三裂叶蟛蜞菊的PSⅡ最大光能转化效率(Fv/Fm)降低了43.8%,而蟛蜞菊只降低了3.7%;同时,与蟛蜞菊相比,三裂叶蟛蜞菊的PSⅡ实际光化学量子产量(Yield)、表观光合电子传递速率(ETR)和光化学猝灭系数(qP)也表现出较大幅度的降低,说明三裂叶蟛蜞菊对干旱胁迫较敏感;但复水后,三裂叶蟛蜞菊能够较快地恢复到正常水平,且与本地种不存在显著差异.这主要是由于入侵种在遭受干旱胁迫时提高了对其过量激发能的热耗散能力以及超氧化物歧化酶(SOD)和抗坏血酸还原酶(APX)的活性,保护光合机构少受不可逆的损伤,使其在干旱胁迫解除后光合功能得以迅速恢复.研究结果初步表明三裂叶蟛蜞菊容易受到水分条件的限制,它向干旱地区扩散的可能性较小.     

Effects of anaerobiosis as probed by the polyphasic chlorophyll a fluorescence rise kinetic in pea (Pisum sativum L.)

We analysed the changes of the chlorophyll (Chl)a fluorescence rise kinetic (from 50 s to 1 s) that occur when leaves or chloroplasts of pea ( Pisum sativum L.) are incubated under anaerobic conditions in the dark. In control leaves, Chl a fluorescence followed a typical O-J-I-P polyphasic rise [Strasser et al. (1995) Photochem Photobiol 61: 32–42]. Anaerobiosis modified the shape of the transient with the main effect being a time-dependent increase in the fluorescence yield at the J-step (2 ms). Upon prolongation of the anaerobic treatment (> 60 min), the O-J-I-P fluorescence rise was eventually transformed to an O-J (J = P) rise. A similar transformation was observed when pea leaves were treated with DCMU or sodium dithionite. Anaerobiosis resulted in a 10–20% reduction in the maximum quantum yield of the primary photochemistry of Photosystem II, as measured by the ratio of the maximal values of variable and total fluorescence (F_V/F_M). When the leaves were returned to the air in the dark, the shape of the fluorescence transient showed a time-dependent recovery from the anaerobiosis-induced change. The original O-J-I-P shape could also be restored by illuminating the anaerobically treated samples with far-red light but not with blue or white light. Osmotically broken chloroplasts displayed under anaerobic conditions fluorescence transients similar to those observed in anaerobically treated leaves, but only when they were incubated in a medium comprising reduced pyridine nucleotides (NADPH or NADH). As in intact leaves, illumination of the anaerobically treated chloroplasts by far-red light restored the original O-J-I-P transient, although only in the presence of methyl viologen. The results provide additional evidence for the existence of a chlororespiratory pathway in higher plant cells. Furthermore, they suggest that the J-level of the fluorescence transient is strongly determined by the redox state of the electron carriers at the PS II acceptor side.     

Nitrate and nitrite as ‘in vivo’ quenchers of chlorophyll fluorescence in blue-green algae

The effect of nitrate and nitrite on long-term chlorophyll fluorescence has been studied in filamentous blue-green algae. Cells grown autotrophically with nitrate as nitrogen source show, under argon atmosphere, a high level of fluorescence. The addition of either nitrete or nitrite induces a significant fluorescence quenching, but, whereas in the case of nitrite no previous treatment is required, in the case of nitrate the cells have to be sonicated or treated with Triton X-100 in advance without destroying their cellular integrity. DCMU strongly inhibits the quenching of fluorescence caused by nitrate or nitrite. Using cells grown with ammonia, a nutritional repressor of the two enzymes of the nitrate-reducing system, the fluorescence quenching observed in either case becomes negligible. These results clearly indicate that both nitrate and nitrite can physiologically act as primary Hill reagents in photosynthesis in blue-green algae.

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Résumé L'effect du nitrate et du nitrite sur la fluorescence de la chlorophylle a été étudié chez quelques algues bleues filameneuses. Les cellules alimentées avec du nitrate montrent, sous argon, un haut niveau de fluorescence. L'addition de quantités égales de nitrate et de nitrite induit une diminution significative de la fluorescence, mais si, dans le cas du nitrite, un traitement préalable n'est pas nécessaire, dans le cas du nitrace, les cellules doivent être traitées légèrement aux ultra sons ou avec du Triton X-100, sans détruire leur intégrité cellulaire. Le DCMU inhibe sévèrement la diminution de fluorescence causée par le nitrate ou le nitrite. Dans les cellules alimentées avec l'ammoniaque, un répresseur des deux enzymes du système de réduction du nitrate, la diminution de fluorescence devient negligeable quel que soit le composé employé. Enfin, ces résultats montrent clairement que tant le nitrate que le nitrite peuvent être des réactifs primaires et physiologiques de la réaction de Hill dans la photosynthèse de nitrate en algues bleues.

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DNA barcoding herbaceous and woody plant species at a subalpine forest dynamics plot in Southwest China

Although DNA barcoding has been widely used to identify plant species composition in temperate and tropical ecosystems, relatively few studies have used DNA barcodes to document both herbaceous and woody components of forest plot. A total of 201 species (72 woody species and 129 herbaceous species) representing 135 genera distributed across 64 families of seed plants were collected in a 25 ha CForBio subalpine forest dynamics plot. In total, 491 specimens were screened for three DNA regions of the chloroplast genome (rbcL, matK, and trnH‐psbA) as well as the internal transcribed spacers (ITS) of nuclear ribosomal DNA. We quantified species resolution for each barcode separately or in combination using a ML tree‐based method. Amplification and sequencing success were highest for rbcL, followed by trnH‐psbA, which performed better than ITS and matK. The rbcL + ITS barcode had slightly higher species resolution rates (88.60%) compared with rbcL + matK (86.60%) and rbcL + trnH‐psbA (86.01%). The addition of trnH‐psbA or ITS to the rbcL + matK barcode only marginally increased species resolution rates, although in combination the four barcodes had the highest discriminatory power (90.21%). The situations where DNA barcodes did not discriminate among species were typically associated with higher numbers of co‐occurring con‐generic species. In addition, herbaceous species were much better resolved than woody species. Our study represents one of the first applications of DNA barcodes in a subalpine forest dynamics plot and contributes to our understanding of patterns of genetic divergence among woody and herbaceous plant species.     

科尔沁沙地22种菊科草本植物叶片形态特征研究

以内蒙古科尔沁沙地22种菊科草本植物为实验材料,研究这些植物的比叶面积、叶片厚度、叶片干物质含量、叶片体积、叶重比和叶片组织密度等特性之间的关系.结果表明,叶片体积和叶片厚度之间存在较强的正相关,其相关系数r=0.782.而叶片厚度与叶片组织密度、比叶面积与叶片组织密度、叶片体积与叶片组织密度之间则存在一定的负相关性,它们的相关系数分别为-0.767、-0.559和-0.505,其余各特性之间均不存在相关性.根据赋值后数据或者二元数据,运用聚类分析方法得出3个组群,并运用单因素方差分析方法检验各特性在各组间的显著性,叶片厚度、叶片体积、叶重比和叶片组织密度特性在组间存在极显著差异.     

Observation and characterisation of a transient in the yield of chlorophyll fluorescence in intact spinach chloroplasts

A transient in chlorophyll fluorescence, which is associated with a transient in 9-aminoacridine fluorescence and a perturbation in the rate of oxygen evolution, has been observed in intact spinach chloroplasts. The results indicate that changes in the redox state of Q are, at least partially, responsible for the transient in chlorophyll fluorescence. The size of the transient is highly dependent upon the concentration of inorganic phosphate and upon the pH of the medium. The properties of the transient are consistent with the suggestion that it reflects changes in the levels of stromal intermediates during induction.Abbreviations BES NN-Bis(2-hydroxyethyl)2-aminoethanesulphonic acid dihydroxyacetone-P(DHAP): dihydroxyacetone phosphate glycerate-3-P (PGA): glycerate-3-phosphate - HEPES N-2-Hydroxyethylpiperazine-N-2-ethanesulphonic acid - MES 2-(N-Morpholino)ethanesulphonic acid - Pi inorganic phosphate - qE quenching of chlorophyll fluorescence by the energisation of the thylakoid membrane - qQ quenching of chlorophyll fluorescence by oxidised Q, the electron acceptor of photosystem 2 - ribose-5-P (R5P) ribose-5-phosphate - Rbu-5-P ribulose-5-phosphate     

Comparison of chlorophyll fluorescence emission characteristics of wheat leaf tissue and isolated thylakoids as a function of excitation wavelength

Abstract. Chlorophyll fluorescence emission spectra and the kinetics of 685 mm fluorescence emission from wheat leaf tissue and thylakoids isolated from such tissue were examined as a function of excitation wavelength. A considerable enhancement of fluorescence emission above 700 nm relative to that at 685 nm was observed from leaf tissue when it was excited with 550 nm rather than 450 nm radiation. Such excitation wavelength dependent changes in the emission spectrum occurred over an excitation spectral range of 440–660 nm and appeared to be directly related to the total quantity of radiation absorbed at a given excitation wavelength. Experiments with isolated thylakoid preparations demonstrated that changes in the fluorescence emission spectrum of the leaf were attributable to the optical properties of the leaf and were not due to the intrinsic characteristies of the thylakoid photochemical apparatus. This was not the case for the observed excitation wavelength dependent changes in the 685 nm fluorescence induction curve obtained from leaf tissue infiltrated with DCMU. Excitation wavelength dependent changes in the ratio of the variable to maximal fluorescence emission and the shape of the variable fluorescence induction were observed for leaf tissue. Isolated thylakoid studies showed that such changes in the leaf fluorescence kinetics were representative of the way in which the photochemical apparatus in vivo was processing the absorbed radiation at the different excitation wavelengths. The results are considered in the context of the use of fluorescence emission characteristics of leaves as non-destructive probes of the photochemical apparatus in vivo.