Leaf characteristics and diurnal variation of chlorophyll fluorescence in leaves of the ‘bana’ vegetation of the amazon region

In six dominant species of the Amazonian ‘Bana’ vegetation, leaf blade characteristics, pigment composition, and chlorophyll (Chl) fluorescence parameters were measured in young and mature leaves under field conditions. Leaf δ¹³C was comparable in the six species, which suggested that both expanding and expanded leaves contained organic matter fixed under similar intercellular and ambient CO₂ concentration (C _i/C _a). High leaf C/N and negative δ¹⁵N values found in this habitat were consistent with the extreme soil N-deficiency. Analysis of Chl and carotenoids showed that expanding leaves had an incomplete development of photosynthetic antenna when compared to adult leaves. Dynamic inactivation of photosystem 2 (PS2) at midday was observed at both leaf ages as F_v/F_m decreased compared to predawn values. Adult leaves reached overnight F_v/F_m ratios typical of healthy leaves. Overnight recovery of F_v/F_m in expanding leaves was incomplete. F₀ remained unchanged from midday to predawn and F_v tended to increase from midday to predawn. The recovery from midday depression observed in adult leaves suggested an acclimatory down-regulation associated with photo-protection and non-damage of PS2.     

Development of photosystems 2 and 1 during leaf growth in grapevine seedlings probed by chlorophyll a fluorescence transient and 820 nm transmission in vivo

Chlorophyll (Chl) a fluorescence transient and 820-nm transmission kinetic were investigated to explore the development of photosynthetic apparatus in grapevine leaves from emergence to full expansion. In this study, all leaves at various developing stages exhibited typical Chl a fluorescence transient. In newly initiating leaves, the maximum quantum yield of primary photochemistry (ϕ_P0) was slightly lower (<10 %) than that in fully expanded leaves. Nevertheless, the fluorescence rise from O to J step was clearly speeded up in young leaves compared with that in fully expanded leaves. Additionally, a distinct K step appeared in young leaves at high irradiances. With leaf development, the efficiency that a trapped exciton can move an electron into the electron transport chain further than Q_A ⁻ (Ψ₀), the quantum yield of electron transport beyond Q_A (ϕ_E0), electron transport flux per excited cross section (ET₀/CS₀), the amount of active photosystem (PS) 2 reaction centres per excited cross section (RC/CS₀), and the performance index on cross section basis (PI_CS) increased gradually and rapidly. Young leaves had strikingly lower amplitude of transmission at 820 nm. A linear relationship between Ψ₀ and the transmission at 820 nm (I₃₀/I₀) was evident. Based on these data, we suggest that (1) the primary photochemistry of PS2 may be not the limiting step of the photosynthetic capacity during leaf growth under natural irradiance; (2) oxygen evolving complex (OEC) might be not fully connected to PS2 at the beginning of leaf growth; (3) though there are a few functional PS1 and PS2 at the early stages of leaf development, they match perfectly.     

Photosynthetic Responses for Vitis vinifera Plants Grown at Different Photon Flux Densities Under Field Conditions

In grapevine (Vitis vinifera L.) leaf chlorophyll (Chl) a and Chl b and carotenoid contents were higher in plants grown at low photon flux densities (PFD) than in those grown at medium and high PFD. The highest Chl a variable to maximum fluorescence ratio F_v/F_m was observed in plants grown at medium PFD while the minimum fluorescence F₀ was highest in those at high PFD. In isolated thylakoids, both high and low PFD caused marked inhibition of whole chain and photosystem 2 (PS2) activities. The artificial exogenous electron donor diphenyl carbazide significantly restored the loss of PS2 activity in low PFD leaves.     

Effect of transient accumulation of anthocyanin on leaf development and photoprotection of Fagopyrum dibotrys mutant

Changes in pigments contents, leaf area, leaf dry mass per unit area (LMA), photosynthetic rate and chlorophyll a fluorescence were investigated in developing leaves of Fagopyrum dibotrys Hara. mutant. Anthocyanins transiently accumulate below the upper epidermis during leaf ontogeny of this mutant. Red leaves possessed lower Chl content, LMA, photosynthetic rate and apparent carboxylation efficiency than green leaves. However, content of anthocyanins declined and above mentioned parameters increased during further leaf development. In both red and green leaves, chronic photoinhibition did not take place according to variable to maximum chlorophyll fluorescence ratio (F_v/F_m). Red leaves had higher non-photochemical quenching (NPQ) and higher PS 2 efficiency.     

Underestimated chlorophyll <Emphasis Type="Italic">a</Emphasis> fluorescence measurements on <Emphasis Type="Italic">Buxus microphylla</Emphasis> red winter leaves

Leaves under stressful conditions usually show downregulated maximum quantum efficiency of photosystem II [inferred from variable to maximum chlorophyll (Chl) a fluorescence (F_v/F_m), usually lower than 0.8], indicating photoinhibition. The usual method to evaluate the degree of photoinhibition in winter red leaves is generally by measuring the F_v/F_m on the red adaxial surface. Two phenotypes of overwintering Buxus microphylla ‘Wintergreen’ red leaves, with different measuring site and leaf thickness, were investigated in order to elucidate how red pigments in the outer leaf layer affected the Chl a fluorescence (F_v/F_m) and photochemical reflectance index. Our results showed that the F_v/F_m measured on leaves with the same red surface, but different leaf thickness, exhibited a slightly lower value in half leaf (separated upper and lower layers of leaves by removing the leaf edge similarly as affected by winter freezing and thawing) than that in the intact leaf (without removing the leaf edge), and the F_v/F_m measured on the red surface was significantly lower than that on the inner or backlighted green surface of the same thickness. Our results suggest that the usual measurement of F_v/F_m on red adaxial surface overestimates the actual degree of photoinhibition compared with that of the whole leaf in the winter.     

The acceptor side of photosystem II is damaged more severely than the donor side of photosystem II in ‘Honeycrisp’ apple leaves with zonal chlorosis

To investigate the photoinhibition of photosynthesis in ‘Honeycrisp’ apple (Malus domestica Borkh. cv. Gala) leaves with zonal chlorosis, we compared pigments, CO₂ assimilation and chlorophyll (Chl) a fluorescence (OJIP) transient between chlorotic leaves and normal ones. Chl and carotenoids (Car) contents, Chl a/b ratio, and absorptance were lower in chlorotic leaves than in normal ones, whereas Car/Chl ratio was higher in the former. Although CO₂ assimilation and stomatal conductance were lower in chlorotic leaves, intercellular CO₂ concentration did not differ significantly between the two leaf types. Compared with normal leaves, chlorotic ones had increased deactivation of oxygen-evolving complexes (OEC), minimum fluorescence (F _o), dissipated energy, relative variable fluorescence at L-, W-, J- and I-steps, and decreased maximum fluorescence (F _m), maximum quantum yield for primary photochemistry (F _v /F _m or TR_o/ABS), quantum yield for electron transport (ET_o/ABS), quantum yield for the reduction of end acceptors of photosystem I (PSI) (φ_Ro and RE_o/ABS), maximum amplitude of IP phase, amount of active photosystem II (PSII) reaction centers (RCs) per cross section (CS) and total performance index (PI_tot,abs). In conclusion, photoinhibition occurs at both the donor (i.e., the OEC) and the acceptor sides of PSII in chlorotic leaves. The acceptor side is damaged more severely than the donor side, which possibly is the consequence of over-reduction of PSII due to the slowdown of Calvin cycle. In addition to decreasing light absorptance by lowering Chl level, energy dissipation is enhanced to protect chlorotic leaves from photo-oxidative damage.     

Photosynthesis in leaves of <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> L. infected with tobacco mosaic virus

In tobacco leaves inoculated with tobacco mosaic virus (TMV), changes in chlorophyll (Chl) and carotenoid contents, parameters of slow Chl fluorescence kinetics, i.e. the maximum quantum yield of photosystem (PS2) photochemistry F_v/F_m, the effective quantum yield of photochemical energy conversion in PS2 Φ₂, ratio of quantum yields of photochemical and concurrent non-photochemical processes in PS2 F_v/F₀, non-photochemical quenching (NPQ), and photochemical activities of isolated chloroplasts from systemically infected tobacco leaves were investigated. We compared two successive stages of infection, the first in the stage of vein clearing at 9^th day post inoculation (dpi) and the second at 22^nd dpi when two different regions, i.e. light- (LGI) or dark-green (DGI) islands in the infected leaf were apparent and symptoms were fully developed. These two different regions were measured separately. The Chl and carotenoid contents in infected leaves decreased with a progression of infection and were lowest in LGI in the second stage. Also the ratio of Chl a/b declined in similar manner. The maximum quantum yield of PS2 photochemistry F_v/F_m, was decreased in the following order: first stage, DGI, and LGI. The same is true for the ratio F_v/F₀. The decrease of Φ₂ in infected leaves declined as compared to their controls. On the contrary, NPQ increased in infected leaves, the highest value was found in the first infection stage. Photochemical activities of the whole electron transport chain in isolated chloroplasts dramatically declined with the progression of symptoms, the lowest value was in LGI. Similarly, but to a lesser extent, the activity of PS2 in isolated chloroplasts decreased in infected leaves. Generally, the most marked impairment of the photosynthetic apparatus was manifested in the LGI of infected leaves.     

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.     

Excess irradiance causes early symptoms of senescence during leaf expansion in photoautotrophically <Emphasis Type="Italic">in vitro</Emphasis> grown tobacco plants

Photosynthetic parameters, growth, and pigment contents were determined during expansion of the fourth leaf of in vitro photoautotrophically cultured Nicotiana tabacum L. plants at three irradiances [photosynthetically active radiation (400–700 nm): low, LI 60 μmol m⁻² s⁻¹; middle, MI 180 μmol m⁻² s⁻¹; and high, HI 270 μmol m⁻² s⁻¹]. During leaf expansion, several symptoms usually accompanying leaf senescence appeared very early in HI and then in MI plants. Symptoms of senescence in developing leaves were: decreasing chlorophyll (Chl) a+b content and Chl a/b ratio, decreasing both maximum (F_V/F_M) and actual (Φ_PS2) photochemical efficiency of photosystem 2, and increasing non-photochemical quenching. Nevertheless, net photosynthetic oxygen evolution rate (P _N) did not decrease consistently with decrease in Chl content, but exhibited a typical ontogenetic course with gradual increase. P _N reached its maximum before full leaf expansion and then tended to decline. Thus excess irradiance during in vitro cultivation did not cause early start of leaf senescence, but impaired photosynthetic performance and Chl content in leaves and changed their typical ontogenetic course.     

Effects of Phytoplasma Infection on Growth and Photosynthesis in Leaves of Field Grown apple (Malus Pumila Mill. cv. Golden Delicious)

The contents of chlorophyll (Chl), leaf biomass, and soluble proteins were markedly decreased in phytoplasma infected apple leaves. Similar results were also observed for ribulose-1,5-bisphosphate carboxylase, ¹⁴CO₂ fixation, and nitrate reductase activity. In contrast, the contents of sugars, starch, amino acids, and total saccharides were significantly increased in phytoplasma infected leaves. In isolated chloroplasts, phytoplasma infection caused marked inhibition of whole photosynthetic electron chain and photosystem 2 (PS2) activity. The artificial exogenous electron donor, diphenyl carbazide, significantly restored the loss of PS2 activity in infected leaves. Similar results were obtained when F_v/F_m was evaluated by in vivo Chl a fluorescence kinetic measurements.     

Effect of gamma radiation on mutant induction of <Emphasis Type="Italic">Fagopyrum dibotrys</Emphasis> Hara

Agronomic traits, photosynthetic pigments, gas exchange, and chlorophyll (Chl) fluorescence parameters of red stem buckwheat (Fagopyrum dibotrys Hara) mutants induced by γ-radiation were compared with green control at seedling stage. Plant height, number of first-class branches, and rhizome biomass were inhibited significantly (p<0.01). Chl a, Chl b, and Chl a+b contents decreased with elevated dose of γ-rays, while increasing carotenoid content indicated that buckwheat was capable of adjusting to the radiation damage. Decrease in net photosynthetic rate was the result of both stomatal and non-stomatal limitations. Fluorescence parameters, such as F₀, F_m, F_v/F_m, F_v/F₀, Φ_PS2, electron transport rate, and photochemical quenching declined significantly (p<0.01) as compared with control due to photoinhibition, while non-photochemical quenching increased to enhance thermal dissipation. Lower parameters implied that leaf tissue was damaged significantly by high dose of γ-radiation and therefore leaf senescence was accelerated.     

Leaf Morphology,Pigment Complex,and Productivity in Wild-Type and afila Pea Genotypes

The effects of genetically determined changes in leaf morphology on the characteristics of growth, pigment complex, and productivity were studied in pea plants (Pisum sativum L.). The homeotic afila(af) mutation, which transformed leaflets into tendrils, decreased the leaf area and the chlorophyll (Chl) content per plant (CCP) in the af/af plants 1.5-fold as compared to the wild type (Af/Af). The loss of leaflets in the af/af plants was partly recompensed by expansion of the tendrils and stipules and by extra accumulation of Chl (a + b). The mutation did not affect Chl (a + b) that fell to the share of light-harvesting complexes (LHC) and the ratio of Chl a/b (representing the relative distribution of chlorophylls between LHC and the reaction centers); neither it affected the quantum efficiency of the primary charge separation (F _v/F _m). The diminished assimilating area (AA) in the af/af plants at the preflowering period did not reduce the final biomass and grain yield. The measurement of the area shaded by plants in the glasshouse experiments and the direct assessment of the vertical profile of solar radiation in the field stand canopies demonstrated that this phenomenon was in particular related to the fact that, in the af/af plants, the solar radiation was available to the apical and subapical leaves (as in the wild-type plants) and also to the lower metamers. As a result, the actively functioning AA expanded, and the photoassimilating potential of the af/af plants was enhanced. Our data presume the direct relationship between plant production and CCP.     

Effect of partial shading on the photosynthetic apparatus and photosystem stoichiometry in sunflower leaves

The partial shading effect on the photosynthetic apparatus of the sunflower (Helianthus annuus L.) was examined by monitoring oxygen evolution, maximum quantum yield of PSII photochemistry in dark-adapted leaves (F_v/F_m), the chlorophyll (Chl) concentrations and the Rubisco contents, and leaf mass per area (LMA) at the leaf level and by determining the concentrations of cytochrome (Cyt) f and the reaction centres of photosystem (PS) I and PSII at the thylakoid level. In this experiment, partial shading was defined as the shading of 2^nd leaves with shade cloths, and the whole treatment was defined as the covering of the whole individuals with shade cloths. In the leaf level responses, oxygen evolution, LMA, Chl concentrations and Rubisco contents decreased in all shade treatments administered for six days. F_v/F_m remained constant irrespective of the shade treatments. On the other hand, in the thylakoid-level responses, the concentrations of the thylakoid components per unit Chl and the stoichiometry of the two photosystems showed no statistical difference among the shade treatments. The data obtained from the present study indicate that the partial shading affected the leaf-level responses rather than the thylakoid-level responses. The light received at the lower leaves might serve as a factor in the regulation of the leaf properties of the upper leaves due to the whole plant photosynthesis, while this factor did not have an effect at the thylakoid level.     

Photosynthesis and protective mechanisms during ageing in transgenic tobacco leaves with over-expressed cytokinin oxidase/dehydrogenase and thus lowered cytokinin content

The content of cytokinins (CKs), the plant inhibitors of the final phase of plant development, senescence, is effectively controlled by irreversible degradation catalysed by cytokinin oxidase/dehydrogenase (CKX). In transgenic tobacco, denoted as AtCKX, with over-expressed CKX causing lowered CK content, we investigated changes in the time courses of chlorophyll (Chl) and xanthophyll (violaxanthin, antheraxanthin, zeaxanthin, neoxanthin, and lutein) contents. We also determined parameters of slow Chl fluorescence kinetics such as minimum Chl fluorescence yield in the darkadapted state F₀, maximum quantum yield of PS2 photochemistry (F_v/F_m), maximum ratio of quantum yields of photochemical and concurrent non-photochemical processes in photosystem 2 (PS2), F_v/F₀, non-photochemical quenching (NPQ), and effective quantum yield of photochemical energy conversion in PS2 (Φ₂). We used three different developmental leaf stages, old, mature, and young, and compared this with time courses of these characteristics in leaves with natural CK levels. The parameters F_v/F_m, F_v/F₀, and Φ₂ were unchanged during ageing in AtCKX plants in contrast to control ones where a significant decrease in old leaves was found. In control plants F₀ increased during ageing, but in the oldest leaf a considerable decrease was observed. This could indicate progressive damage to PS2 reaction centres and then detachment and rapid degradation of Chl. This is in agreement with time course of Chl content. NPQ decreased with age and was similar in both plant types. We observed a decline of xanthophyll contents in the oldest leaves in both plant types, but the contents were enhanced in AtCKX compared to control plants, especially of neoxanthin. The higher xanthophyll contents in the transgenic plants contribute to a better photoprotection and the fluorescence parameters indicated that photosynthetic apparatus was in better condition compared to control and it consequently postponed the onset of leaf senescence.     

Response of photosynthesis and chlorophyll fluorescence quenching to leaf dichotocarpism in Ligustrum vicaryi, an ornamental herb

Net photosynthetic rate of yellow upper leaves (UL) of Ligustrum vicaryi was slightly, but not significantly higher than that of green lower leaves (LL). Diurnally, maximum photochemical efficiency of photosystem 2, PS2 (F_v/F_m) of LL did not significantly decline but the UL showed fairly great daily variations. Yield of PS2 of UL showed an enantiomorphous variation to the photosynthetically active radiation and was significantly lower than in the LL. Unlike F_v/F_m, the efficiency of energy conversion in PS2 and both non-photosynthetic and photosynthetic quenching did not differ in UL and LL. Significant differences between UL and LL were found in contents of chlorophyll (Chl) a, b, and carotenoids (Car) and ratios of Chl a/b, Chl b/Chl (a+b), and Car/Chl (a+b). Leaf colour dichotocarpism in L. vicaryi was mainly caused by different photon utilization; sunflecks affected the LL.     

Changes in stomatal characteristics and photochemical efficiency during leaf development in six species of Sorbus

Measurements of Sorbus stomata size and density, maximal photochemical efficiency of photosystem II (F_v/F_m), variable-to-initial fluorescence ratio (F_v/F₀) and potential electron acceptor capacity (‘area’) were performed during leaf development in four parental diploid species, S. aria, S. aucuparia, S. chamaemespilus, S. torminalis, and two hybrid species, S. hazslinszkyana and S. intermedia. In fully expanded mature leaves, stomata lengths and densities were significantly larger in the shrub S. chamaemespilus than in the five tree species. The best performance of both the F_v/F_m and the F_v/F₀ ratio was recorded in S. intermedia, whereas S. chamaemespilus had the highest value of ‘area’. From a physiological point of view, the results of this study showed that the photosystem II reaction centers remained intact functionally through all phenological stages of leaf expansion for all examined species of Sorbus.     

Photosynthetic activity of poikilochlorophyllous desiccation tolerant plant <Emphasis Type="Italic">Reaumuria soongorica</Emphasis> during dehydration and re-hydration

Diurnal patterns of gas exchange and chlorophyll (Chl) fluorescence parameters of photosystem 2 (PS2) as well as Chl content were analyzed in Reaumuria soongorica (Pall.) Maxim., a perennial semi-shrub during dehydration and rehydration. The net photosynthetic rate (P _N), maximum photochemical efficiency of PS2 (variable to maximum fluorescence ratio, F_v/F_m), quantum efficiency of non-cyclic electron transport of PS2, and Chl content decreased, but non-photochemical quenching of fluorescence and carotenoid content increased in stems with the increasing of drought stress. 6 d after re-hydration, new leaves budded from stems. In the re-watered plants, the chloroplast function was restored and Chl a fluorescence returned to a similar level as in the control plants. This improved hydraulic adjustment in plant triggered a positive effect on ion flow in the tissues and increased shoot electrical admittance. Thus R. soongorica plants are able to sustain drought stress through leaf abscission and keep part of Chl content in stems.     

Chlorophyll fluorescence imaging of photosynthetic activity with the flash-lamp fluorescence imaging system

A flash-lamp chlorophyll (Chl) fluorescence imaging system (FL-FIS) is described that allows to screen and image the photosynthetic activity of several thousand leaf points (pixels) of intact leaves in a non-destructive way within a few seconds. This includes also the registration of several thousand leaf point images of the four natural fluorescence bands of plants in the blue (440 nm) and green (520 nm) regions as well as the red (near 690 nm) and far-red (near 740 nm) Chl fluorescence. The latest components of this Karlsruhe FL-FIS are presented as well as its advantage as compared to the classical single leaf point measurements where only the fluorescence information of one leaf point is sensed per each measurement. Moreover, using the conventional He-Ne-laser induced two-wavelengths Chl fluorometer LITWaF, we demonstrated that the photosynthetic activity of leaves can be determined measuring the Chl fluorescence decrease ratio, R_Fd (defined as Chl fluorescence decrease F_d from maximum to steady state fluorescence F_s:F_d/F_s), that is determined by the Chl fluorescence induction kinetics (Kautsky effect). The height of the values of the Chl fluorescence decrease ratio R_Fd is linearly correlated to the net photosynthetic CO₂ fixation rate P _N as is indicated here for sun and shade leaves of various trees that considerably differ in their P _N. Imaging the R_Fd-ratio of intact leaves permitted the detection of considerable gradients in photosynthetic capacity across the leaf area as well as the spatial heterogeneity and patchiness of photosynthetic quantum conversion within the control leaf and the stressed plants. The higher photosynthetic capacity of sun versus shade leaves was screened by Chl fluorescence imaging. Profile analysis of fluoresence signals (along a line across the leaf area) and histograms (the signal frequency distribution of the fluorescence information of all measured leaf pixels) of Chl fluorescence yield and Chl fluorescence ratios allow, with a high statistical significance, the quantification of the differences in photosynthetic activity between various areas of the leaf as well as between control leaves and water stressed leaves. The progressive uptake and transfer of the herbicide diuron via the petiole into the leaf of an intact plant and the concomitant loss of photosynthetic quantum conversion was followed with high precision by imaging the increase of the red Chl fluorescence F₆₉₀. Differences in the availability and absorption of soil nitrogen of crop plants can be documented via this flash-lamp fluorescence imaging technique by imaging the blue/red ratio image F₄₄₀/F₆₉₀, whereas differences in Chl content are detected by collecting images of the fluorescence ratio red/far-red, F₆₉₀/F₇₄₀.     

Changes in stomatal frequency, stomatal conductance and cuticle thickness during leaf expansion in the broad-leaved evergreen species, Eucalyptus regnans

Aspects of leaf anatomical and physiological development were investigated in the broad-leaved evergreen species, Eucalyptus regnans F.Muell. Newly emergent leaves were tagged in the field and measured for stomatal conductance while a subset was collected every 14 days for the measurement of stomata and cuticle over a 113-day period. Cuticle thickness increased during leaf expansion, the increase following a sigmoid curve. Stomatal frequency (no. mm⁻²) decreased from 56 to 113 days after leaf emergence. The frequency of both immature and intermediate developmental stages of stomata also decreased over this time, but the total number of stomata per leaf remained relatively constant. Stomatal conductance (g _s) of young expanding leaves increased during expansion, and was significantly linearly correlated with stomatal frequency (excluding immature stomata), and with cuticle thickness. The progressive increase in g _s in young developing leaves was contrary to the observed changes in structural characteristics (increased cuticle thickness and decreased stomatal frequency). This increase in g _s with development may be related to the progressive increase in number of mature stomata with larger apertures and, therefore, a higher total pore area in fully expanded leaves.     

Rehydration of Sugar Beet Plants after Water Stress: Effect of Cytokinins

The possibility to improve the recovery of sugar beet plants after water stress by application of synthetic cytokinins N⁶-benzyladenine (BA) or N⁶-(m-hydroxybenzyl)adenosine (HBA) was tested. Relative water content (RWC), net photosynthetic rate (P_N), transpiration rate (E), stomatal conductance (g_s), chlorophyll (Chl) a and Chl b contents, and photosystem 2 efficiency characterized by variable to maximal fluorescence ratio (F_v/F_m) were measured in control plants, in water-stressed plants, and after rehydration (4, 8, 24, and 48 h). Water stress markedly decreased parameters of gas exchange, but they started to recover soon after irrigation. Application of BA or HBA to the substrate or sprayed on leaves only slightly stimulated recovery of P_N, E, and g_s in rehydrated plants, especially during the first phases of recovery. Chl contents decreased only under severe water stress and F_v/F_m ratio was not significantly affected by water stress applied. Positive effects of BA or HBA application on Chl content and F_v/F_m ratio were mostly not observed.