How to correctly determine the different chlorophyll fluorescence parameters and the chlorophyll fluorescence decrease ratio R<Subscript>Fd</Subscript> of leaves with the PAM fluorometer

This contribution is a practical guide to the measurement of the different chlorophyll (Chl) fluorescence parameters and gives examples of their development under high-irradiance stress. From the Chl fluorescence induction kinetics upon irradiation of dark-adapted leaves, measured with the PAM fluorometer, various Chl fluorescence parameters, ratios, and quenching coefficients can be determined, which provide information on the functionality of the photosystem 2 (PS2) and the photosynthetic apparatus. These are the parameters F_v, F_m, F₀, F_m′, F_v′, NF, and ΔF, the Chl fluorescence ratios F_v/F_m, F_v/F₀, ΔF/F_m′, as well as the photochemical (q_P) and non-photochemical quenching coefficients (q_N, q_CN, and NPQ). q_N consists of three components (q_N = q_E + q_T + q_I), the contribution of which can be determined via Chl fluorescence relaxation kinetics measured in the dark period after the induction kinetics. The above Chl fluorescence parameters and ratios, many of which are measured in the dark-adapted state of leaves, primarily provide information on the functionality of PS2. In fully developed green and dark-green leaves these Chl fluorescence parameters, measured at the upper adaxial leaf side, only reflect the Chl fluorescence of a small portion of the leaf chloroplasts of the green palisade parenchyma cells at the upper outer leaf half. Thus, PAM fluorometer measurements have to be performed at both leaf sides to obtain information on all chloroplasts of the whole leaf. Combined high irradiance (HI) and heat stress, applied at the upper leaf side, strongly reduced the quantum yield of the photochemical energy conversion at the upper leaf half to nearly zero, whereas the Chl fluorescence signals measured at the lower leaf side were not or only little affected. During this HL-stress treatment, q_N, q_CN, and NPQ increased in both leaf sides, but to a much higher extent at the lower compared to the upper leaf side. q_N was the best indicator for non-photochemical quenching even during a stronger HL-stress, whereas q_CN and NPQ decreased with progressive stress even though non-photochemical quenching still continued. It is strongly recommended to determine, in addition to the classical fluorescence parameters, via the PAM fluorometer also the Chl fluorescence decrease ratio R_Fd (F_d/F_s), which, when measured at saturation irradiance is directly correlated to the net CO₂ assimilation rate (_{P N}) of leaves. This R_Fd-ratio can be determined from the Chl fluorescence induction kinetics measured with the PAM fluorometer using continuous saturating light (cSL) during 4–5 min. As the R_Fd-values are fast measurable indicators correlating with the photosynthetic activity of whole leaves, they should always be determined via the PAM fluorometer parallel to the other Chl fluorescence coefficients and ratios.     

Facilitated water transport in cyanobacterium Synechococcus sp. PCC 7942 studied by phycobilisome-sensitized chlorophyll a fluorescence

Water transport across plant cell membranes is difficult to measure. We present here a model assay, based on chlorophyll (Chl) a fluorometry, with which net water transport across the cell membrane of freshwater cyanobacterium Synechococcus sp. PCC7942 (S7942) can be followed kinetically with millisecond-time resolution. In cyanobacteria, the phycobilisome (PBS)-sensitized Chl a fluorescence increases when cells expand (e.g., in hypo-osmotic suspension) and decreases when cells contract (e.g., in hyper-osmotic suspension). The osmotically-induced Chl a fluorescence changes are proportional to the reciprocal of the suspension osmolality (ΔF ∝ Osm⁻¹; Papageorgiou GC and Alygizaki-Zorba A (1997) Biochim Biophys Acta 1335: 1–4). In our model assay, S7942 cells were loaded with NaCl (passively penetrating solute) and shrunk in hyper-osmotic glycine betaine (nonpenetrating solute). Upon injecting these cells into hypo-osmotic medium, the PBS-sensitized Chl a fluorescence rose to a maximum due to the osmotically-driven water uptake. The rise of Chl a fluorescence (water uptake) was partially inhibited by HgCl₂, at micromolar concentrations. Arrhenius plots of the water uptake rates gave activation energies of E_A=4.9 kcal mol⁻¹, in the absence of HgCl₂, and E_A=11.9 kcal mol⁻¹ in its presence. These results satisfy the usual criteria for facilitated water transport through protein water pores of plasma membranes (aquaporins), namely sensitivity to Hg²⁺ ions and low activation energy.     

不同微生境中水光温变化对毛尖紫萼藓叶绿素荧光特性的影响

毛尖紫萼藓（Grimmia pilifera）多生长在裸岩表面且具有多种微生境,其生长过程受到水分、光照和温度等环境因素的交互影响,但其光合生理特征如何响应这种变化的环境条件尚不清楚。开展原位（荫蔽和向阳裸岩2种微生境）和室内模拟实验,分析了不同水分（模拟降水量和降水频次）、光照、温度及其复合梯度处理对毛尖紫萼藓叶绿素荧光特性的影响。原位实验结果显示荫蔽生境原位生长毛尖紫萼藓光化学效率显著高于向阳生境。在室内相同培养条件下,脱水过程中来自荫蔽生境植株有效光合效率保持时间比向阳生境明显缩短,表现出较低的脱水耐受性。水-光-温复合模拟实验显示,降水频次和光-温变化对毛尖紫萼藓光化学效率均有极显著影响且存在一定的交互作用,而降水量的影响相对较弱;弱光低温及1次/（2 d）的降水频率条件下毛尖紫萼藓具有最高的光合活性。总体来看,荫蔽生境、弱光低温条件及中等频次降水有利于毛尖紫萼藓生长,但向阳生境毛尖紫萼藓则具有更强的环境耐受性。     

水体铜污染对苦草生长及叶绿素荧光特性的影响

该文以苦草(Vallisneria natans)为材料,研究了不同浓度铜污染水体对其叶/根长、生物量、光合色素含量、体内重金属含量及叶绿素荧光参数的影响。结果表明:随着水体铜浓度的增加,苦草的叶长、根长、生物量均显著下降;光合色素含量逐渐下降,其中叶绿素a比叶绿素b下降明显,类胡萝卜素下降幅度最小;叶片铜含量随铜浓度的增加显著上升(P0.05),各处理组根铜含量没有显著性差异,在水中铜低于0.5mg·L-1的环境中苦草具有正常的光合活性。除0.5 mg·L-1处理组外,最大荧光(Fm)、最大光化学效率(Fv/Fm)、潜在光化学效率(Fv/Fo)、光化学淬灭系数(qP)与有效量子产量[Y(Ⅱ)]均比对照组显著降低(P0.05),而非光化学淬灭效率(qN)、调节性能量耗散的量子产量[Y(NPQ)]、非调节性能量耗散的量子产量[Y(NO)]均呈上升趋势,其中Fv/Fo对铜污染反应最为灵敏。综合各参数变化情况,随着水体铜浓度的增加,苦草生长受到抑制,叶片利用光能的效率下降,PSⅡ反应中心的电子传递受到明显的抑制;苦草通过自身调节以热的形式将过剩光能耗散,以减轻PSⅡ反应中心受伤害的程度;苦草是铜超富集植物,其可作为低浓度铜污染水体生态修复的备选物种。     

Sensitivity of the Algal Biotest ISO 10253 to the Photosystem 2 Herbicides in Seawater

The sensitivity of marine algal biotest ISO 10253 to the photosystem 2 (PS2) herbicide diuron (DCMU) was determined. Using the diatom Phaeodactylum tricornutum, we found that the algal growth rate was reduced to 50 % of the control value (EC₅₀) for ca. 200 nM DCMU. This value is too high to allow a practical application of the biotest for concentrations of the PS2 herbicides found in natural waters. The mechanisms causing the low sensitivity of the biotest to the PS2 herbicide were investigated by measuring parameters of photosynthetic apparatus in the diatom prior and during the biotest. The apparent dissociation constant for DCMU in P. tricornutum found by measurements of inhibition of oxygen evolution and of variable fluorescence was in the range 60–90 nM. This should lead to a much higher sensitivity of the biotest than found in our experiments. The low biotest sensitivity is caused by an acclimation to sub-lethal DCMU concentrations. The acclimation is manifested by the chlorophyll content per cell that is increasing with the DCMU concentration. During a prolonged exposure to sub-lethal herbicide concentrations, we observed also a selection of DCMU resistant organisms indicating that also an adaptation may decrease the test sensitivity. The biotest sensitivity may increase when the acclimation and adaptation are limited by shortening of the experiment duration. This revised version was published online in August 2006 with corrections to the Cover Date.     

Interaction of water quantity with water quality: the Lake Chapala example

Water quality may be significantly determined by water quantity. Lake Chapala, México is a large lake beset with numerous water quality problems. The decline in water volume over the past 20 years, a serious problem itself, is associated with causing or enhancing several problems of quality. Five such problems are explored herein. These are: extensive infestations of water hyacinth (Eichhornia crassipes), a declining native fishery, light limitation of phytoplankton production at the base of the food chain, shallow-water algal blooms resulting in water supply treatment problems, and the presence of toxic metals in the harvested and sold fishes.     

The use of chlorophyll fluorescence as a screening method for cold tolerance in maize

Chlorosis in maize (Zea mays L.) is a common phenomenon in the 12 to 17°C temperature range. A newly developed chlorophyll-fluorescence technique was used to elucidate the underlying subcellular processes of resistance to chlorosis. Four populations were used that were developed by divergent mass selection for contrasting resistance to chlorosis in a cold-tolerant dent and a cold-tolerant flint population. Young plants from the four populations were kept for six days at 17/10, 15/10 and 13/10°C (day/night). After 1, 3 and 6 days various chlorophyll-fluorescence parameters were determined. The measurements were done on leaf 4. Differences were not uniform for all fluorescence properties. The resistant and susceptible populations of the two sets differed for the Q-quenching which is related to the electron transport rate in the chloroplast. For the E-quenching which is related to the Calvin cycle activity, the resistant dent differed significantly from the other three populations. The ratio Fm/Fo (related to the transfer of absorbed light-energy from antennae pigments to reaction centers in the chloroplast) was higher for the resistant dent population than for the susceptible one. The flint types did not differ for this property.Apparently, divergent mass selection for chlorosis resistance resulted in various changes at the subcellular level that are not necessarily comparable for flint and dent types.When after 6 days the temperature was raised from 13°C to 17°C, the fluorescence signals led to the conclusion that there was a full recovery of various processes after two days, except for the metabolic activity of the susceptible flint.     

The effect of NaCl on water relations,chlorophyll, and protein and proline contents of two cultivars of blackgram (Vigna mungo L.)

The physiological basis of salt tolerance of two cultivars of blackgram, cv Candhari Mash (relatively salt tolerant) and cv Mash 654 (salt sensitive), was assessed in salinized sand culture at the flowering stage. Increasing NaCl concentration in the rooting medium significantly reduced the chlorophyll a, chlorophyll b, and total chlorophyll, leaf water potential (Ψ_w), leaf solute potential (Ψ_s), and leaf turgor potential (Ψ_p) in both the cultivars. Leaf protein and proline content was increased as a result of increasing salt concentration in both cultivars. High salt concentrations had no significant effect on the seed protein content of both cultivars. At high salinities, cv Candhari Mash had significantly greater chlorophyll a, chlorophyll b and total chlorophyll, leaf water potential, solute potential, and turgor potential than cv Mash 654, but the latter had greater leaf proline content than cv Candhari Mash. Cultivars did not differ significantly for both leaf and seed protein contents. The relatively salt tolerant cv Candhari Mash maintained high leaf water potential and turgor potential to resist salt injury. Leaf proline content had negative correlation with salt tolerance in blackgram.     

水分胁迫与复水对地枫皮生理生态特性的影响

以岩溶特有药用植物地枫皮为材料,研究土壤水分胁迫及复水条件下,其叶片光合参数、叶绿素荧光参数及光合色素含量的变化特性,进而探讨其对水分胁迫的生理生态适应性。结果表明:停止供水10 d,水分胁迫地枫皮叶片的P_n(净光合速率)、C_i(胞间CO_2浓度)、G_s(气孔导度)和L_s(气孔限制值)均下降,气孔限制是P_n降低的主要原因;停止供水15 d,水分胁迫地枫皮叶片的P_n日变化呈逐渐下降趋势,上午9:30以后全天的P_n值均接近零,非气孔限制成为P_n下降的主要因素;而对照地枫皮叶片的P_n日变化呈"双峰型",中午P_n下降的主要原因依然是气孔限制。水分胁迫下,地枫皮叶片叶绿素含量降低和Chl_(a/b)升高,减少了叶片对光能的捕获,减轻了光合机构遭受光氧化的破坏,而Car/Chl_(a+b)升高增强了光保护能力。水分胁迫下,地枫皮叶片的初始荧光(F_o)显著增大,最大荧光(F_m)、光系统Ⅱ(PSⅡ)潜在活性(F_v/F_o)和最大光化学效率(F_v/F_m)均显著降低,表明水分胁迫对地枫皮叶片的PSⅡ反应中心和电子传递造成了一定的破坏,从而使其PSⅡ的潜在活性和最大光化学效率降低。复水5 d后,地枫皮的上述生理生态参数均能恢复到对照水平,表明其复水后的生理修复能力很强。     

Light-harvesting chlorophyll a-b complex requirement for regulation of Photosystem II photochemistry by non-photochemical quenching

Recently, it has been suggested (Horton et al. 1992) that aggregation of the light-harvesting a-b complex (LHC II) in vitro reflects the processes which occur in vivo during fluorescence induction and related to the major non-photochemical quenching (qE). Therefore the requirement of this chlorophyll a-b containing protein complex to produce qN was investigated by comparison of two barley mutants either lacking (chlorina f2) or depressed (chlorina¹⁰⁴) in LHC II to the wild-type and pea leaves submitted to intermittent light (IL) and during their greening in continuous light. It was observed that qN was photoinduced in the absence of LHC II, i.e. in IL grown pea leaves and the barley mutants. Nevertheless, in these leaves qN had no (IL, peas) or little (barley mutants) inhibitory effect on the photochemical efficiency of Q_A reduction measured by flash dosage response curves of the chlorophyll fluorescence yield increase induced by a single turn-over flash During greening in continuous light of IL pea leaves, an inhibitory effect on Q_A photoreduction associated to qN developed as Photosystem II antenna size increased with LHC II synthesis. Utilizing data from the literature on connectivity between PS II units versus antenna size, the following hypothesis is put forward to explain the results summarized above. qN can occur in the core antenna or Reaction Center of a fraction of PS II units and these units will not exhibit variable fluorescence. Other PS II units are quenched indirectly through PS II-PS II exciton transfer which develops as the proportion of connected PS II units increases through LHC II synthesis.     

Photosynthetic parameters and leaf water potential of five common bean genotypes under mild water deficit

The leaf water potential, gas exchange and chlorophyll fluorescence were evaluated in five common bean (Phaseolus vulgaris) genotypes A222, A320, BAT477, Carioca and Ouro Negro subjected to moderate water deficit. At the maximum water deficit (10 d of water withholding), the leaf water potential of genotypes A320 and A222 was higher (−0.35 and −0.50 MPa) when compared to the other genotypes (−0.67 to −0.77 MPa). The stomatal conductance and net photosynthetic rate were significantly reduced in all genotypes due to the water deficit. The greater reduction in stomatal conductance of A320 under drought resulted in high intrinsic water use efficiency. Mild water deficit affected the photochemical apparatus in bean genotypes probably by down-regulation since plants did not show photoinhibition. The photochemical apparatus of A222 and A320 genotypes was more sensitive to drought stress, showing reduced apparent electron transport even after the recovery of plant water status. On the other hand, even after 10 d of water withholding, the maximum efficiency of photosystem 2 was not affected, what suggest efficiency of the photoprotection mechanisms.     

不同油松种源光合和荧光参数对水分胁迫的响应特征

采用盆栽控水试验方法,设置正常水分(T1,田间持水量的70％-80％)、轻度胁迫(T2,田间持水量的50％-60％)和严重胁迫(T3,田间持水量的30％-40％)3个水分梯度,对6个油松(Pinus tabulaeformis Carr.)种源(陕西洛南LN,陕西桥山 QO,山西灵空山LK,辽宁千山QN,河北雾灵山WL和山西芦芽山LY)的光合参数和叶绿素荧光参数进行比较,探讨种源间光合和水分利用特征的差异及其与气孔导度和荧光参数的关系.结果表明,3个水分梯度下6个种源的净光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)、水分利用效率(WUE)、原初光能转换效率(Fv/Fm)和PSⅡ潜在活性(Fv/Fo)差异均极显著(P＜0.01),在T1和T3下,各种源的实际光量子效率φPSⅡ和电子传递速率ETR差异显著(P＜0.05).在T2下,LK和WL的Pn达到最大,具有轻度胁迫下提高光合生产力的倾向.在T3下,QN种源的Pn和WUE较高(除LK的WUE最高外),而WL的Pn和WUE较低(除LK的Pn最低外);6个种源中QN的Pn、φPSⅡ和ETR最大,LY和QO的光合生产力仅次于QN,而LN、LK和WL的Pn、Gs、φPSⅡ和ETR比较低,其光合生产力及光量子产量受干旱胁迫影响较大.LK的WUE在各个水分梯度下均显著高于其它种源(P＜0.05).在T3下,各种源通过降低Tr提高WUE.在T1下,气孔限制是影响油松Pn的主要因素;在T2和T3下,Pn和WUE均与气孔导度、PSⅡ反应中心的光量子产量和电子传递速率密切相关.     

Use of simultaneous analysis of gas-exchange and chlorophyll fluorescence quenching for analysing the effects of water stress on photosynthesis in apple leaves

Summary A convenient system for the rapid simultaneous measurement of both chlorophyll fluorescence quenching using a modulated light system, and of CO₂, and water vapour exchange by leaves is described. The system was used in a study of the effects of water deficits on the photosynthesis by apple leaves (Malus x domestica Borkh.). Apple leaves were found to have low values of steady-state variable fluorescence, and the existence of significant fluorescence with open traps (F_o) quenching necessitated the measurement and use of a corrected F_o in the calculation of quenching components. Long-term water stress had a marked effect on both gas-exchange and chlorophyll fluorescence quenching. Non-photochemical quenching (qn) in particular was increased in water-stressed leaves, and it was particularly sensitive to incident radiation in such leaves. In contrast, rapid dehydration only affected gas exchange. Relaxation of qn quenching in the dark was slow, taking approximately 10 min for a 50% recovery, in well-watered and in draughted plants, and whether or not the plants had been exposed to high light.     

水分胁迫对极小种群东兴金花茶幼苗光合特性的影响

为了解东兴金花茶幼苗对水分胁迫的适应能力和响应机制,该文以东兴金花茶1年生实生苗为材料,采用盆栽控水试验,研究不同控水时间处理对东兴金花茶幼苗的生理生态特性的影响。结果表明:随着控水时间的延长,水分胁迫的程度不断加剧,东兴金花茶叶片净光合速率(P_n)、气孔导度(G_s)、蒸腾速率(T_r)呈现显著下降趋势;胞间CO₂浓度(C_i)呈现先低后高的变化趋势,水分利用效率(WUE)呈现先高后低的变化趋势。土壤含水率和叶片相对含水量均呈现不断下降的趋势,丙二醛呈现先降后升的变化趋势;东兴金花茶幼苗的荧光参数F_v/F_m和F_v/F_o呈现先增加后不断下降趋势,分别从0.806下降至0.754和4.17下降至3.08,表明水分胁迫降低了PSⅡ原初光能转化效率,光合作用原初反应过程受到抑制。基于水分胁迫的生理生态指标和叶片生物性状的变化表明,控水时间在4 d情况下东兴金花茶可以提高自身水分利用效率来抵抗干旱,说明东...     

The influence of mineral salts on fecundity of the water flea (Daphnia magna) and the implications on toxicity testing of industrial wastewater

The effects of mineral salts constituting water hardness on fecundity ofDaphnia magna were assessed. Of the salts tested, increased concentrations of NaHCO₃ and MgSO₄ had no effect on fecundity, CaSO₄ significantly increased fecundity, and KCl significantly reduced fecundity. The number of offspring produced per daphnid was correlative to the CaSO₄ concentration at CaSO₄ concentrations between 91 and 2100 mg/ℓ. The effects of CaSO₄ on daphnid fecundity could influence the interpretive outcome of industrial wastewater toxicity tests using this species when the waste and dilution waters contain different concentrations of CaSO₄. It is recommended that when performing these tests, dilution water be sampled at the intake site of the industry's water source, thus assuring initial comparability of the waste and dilution waters. The CaSO₄ content of the water prior to and after industrial use should be determined to identify any alterations of CaSO₄ concentration during use. Identification of CaSO₄ concentration differences can aid in the interpretation of effects associated with the wastewater.     

Removal of caffeine in sewage by Pseudomonas putida: Implications for water pollution index

A strain of Pseudomonas putida (biotype A) capable of growing on caffeine (1,3,7-trimethylxanthine) was isolated from a domestic wastewater processing operation. It used caffeine as the sole carbon source with a mean growth rate constant (k) of 0.049 h^-1 (approximately 20 h per generation), whereas k for glucose utilization under similar incubation conditions was 0.31 (3.3 h per generation). The isolate contained at least two plasmids, and the increased expression of a 40 kDa protein was attributable to growth on caffeine. Degradation byproducts of caffeine metabolism by the bacterial isolate included other xanthine derivatives. The slow bacterial catabolism of caffeine in sewage has implications for the effectiveness of wastewater purification, re-use and disposal.The author is with the Laboratory for Molecular Ecology, Department of Environmental Analysis and Design, University of California at Irvine, Irvine, CA 92717-5150 U.S.A.     

Application of the microalga Plagioselmis prolonga for the assessment of water quality in the Amursky and Nakhodka Bays (Sea of Japan)

Based on biotesting, we carried out an estimation of the water quality in the Amursky and Nakhodka Bays (Sea of Japan) using Plagioselmis prolonga (Cryptophyta). The obtained data were compared with the data from water biotesting using Dunaliella salina (Chlorophyta). It was shown that water from the Amursky Bay produced more a negative effect on both microalgae than water from the Nakhodka Bay. It was established that sensitivity of the P. prolonga microalga exceeded that of D. salina. This was confirmed by a sharp decrease of the P. prolonga motile cell number in the studied water.     

Aquatic plant bioassays used in the assessment of water quality in German rivers

A novel aquatic bioassay with Lemna minor (duckweed), a representative of higher plants, was applied to pore waters (interstitial waters) from river sediments. The results have been used for the ecotoxicological assessment of sediments in several German rivers. The aim of the present study is to demonstrate the usefulness of a macrophyte bioassay, as a supplement to the standardised algae bioassays, with regard to the ecotoxicological assessment of aquatic systems. Results relating to the development of a sediment contact test (using the whole sediment) with Lemna minor are also presented.     

Influence of water stress on leaf photosynthetic characteristics in wheat cultivars differing in their susceptibility to drought

A gradual reduction in leaf water potential (Ψ_leaf), net photosynthetic rate (P _N), stomatal conductance, and transpiration rate was observed in two drought tolerant (C 306 and K 8027) and two susceptible (RW 893 and 899) genotypes subjected to water stress. The extent of reduction was lower in K 8027 and C 306 and higher in RW 893 and RW 899. Rewatering the plants after 5 d of stress restored P _N and other gas exchange traits in all four cultivars. Water stress had no significant effect on variable to maximum fluorescence ratio (F_v/F_m) indicating that water stress had no effect on primary photochemistry of photosystem 2 (PS2). However, water stress reduced the efficiency of excitation energy transfer (F′_v/F′_m) and the quantum yield of electron transport (Φ_PS2). The reduction was more pronounced in susceptible cultivars. Water stress had no significant effect on photochemical quenching, however, the non-photochemical quenching increased by water stress.     

New Fluorescence Parameters for the Determination of QA Redox State and Excitation Energy Fluxes

A number of useful photosynthetic parameters are commonly derived from saturation pulse-induced fluorescence analysis. We show, that q(P), an estimate of the fraction of open centers, is based on a pure 'puddle' antenna model, where each Photosystem (PS) II center possesses its own independent antenna system. This parameter is incompatible with more realistic models of the photosynthetic unit, where reaction centers are connected by shared antenna, that is, the so-called 'lake' or 'connected units' models. We thus introduce a new parameter, q(L), based on a Stern-Volmer approach using a lake model, which estimates the fraction of open PS II centers. We suggest that q(L) should be a useful parameter for terrestrial plants consistent with a high connectivity of PS II units, whereas some marine species with distinct antenna architecture, may require the use of more complex parameters based on intermediate models of the photosynthetic unit. Another useful parameter calculated from fluorescence analysis is Phi(II), the yield of PS II. In contrast to q(L), we show that the Phi(II) parameter can be derived from either a pure 'lake' or pure 'puddle' model, and is thus likely to be a robust parameter. The energy absorbed by PS II is divided between the fraction used in photochemistry, Phi(II), and that lost non-photochemically. We introduce two additional parameters that can be used to estimate the flux of excitation energy into competing non-photochemical pathways, the yield induced by downregulatory processes, Phi(NPQ), and the yield for other energy losses, Phi(NO).