ALBERT

Notizen: The profile of photosynthetic rate with depth through a leaf depends on the profiles of light absorption and photosynthetic capacity. Using a combination of several techniques, a comprehensive description of spinach leaves has been obtained. Profiles of CO2 fixation were obtained by exposing leaves to 14CO2 for 10 s under blue or green light before freeze clamping and paradermal sectioning. Profiles of light absorption were measured on adjacent parts of the leaf by quantifying chlorophyll fluorescence images of the transversely cut face obtained when blue or green light was applied to the adaxial or abaxial surface. The profile of CO2 fixation was modelled using the measured profiles of light absorption and photosynthetic capacity. There was remarkably good agreement between the observed and modelled CO2 fixation profiles for the eight combinations of colour, orientation and irradiance tested. Gas exchange of an intact leaf was also measured concurrently with conventional chlorophyll fluorescence under blue or green light. These data were consistent with the multi-layer leaf model with the exception of blue light applied to the abaxial surface, where chlorophyll fluorescence appeared to come from layers deeper than expected. Photosynthetic capacity matched the profile of green but not white light absorption through the leaf.

Notizen: Chlorophyll fluorescence was used to estimate profiles of absorbed light within chlorophyll solutions and leaves. For chlorophyll solutions, the intensity of the emitted fluorescence declined in a log–linear manner with the distance from  the  irradiated  surface  as  predicted  by  Beer's  law. The amount of fluorescence was proportional to chlorophyll  concentration  for  chlorophyll  solutions  given  epi-illumination on a microscope slide. These relationships appeared to hold for more optically complex spinach leaves. The profile of chlorophyll fluorescence emitted by leaf cross sections given epi-illumination corresponded to chlorophyll content measured in extracts of leaf paradermal sections. Thus epifluorescence was used to estimate relative chlorophyll content through leaf tissues. Fluorescence profiles across leaves depended on wavelength and orientation, reaching a peak at 50–70 µm depth. By infiltrating leaves with water, the pathlengthening due to scattering at the airspace : cell wall interfaces was calculated. Surprisingly, the palisade and spongy mesophyll had similar values for pathlengthening with the value being greatest for green light (550 〉 650 〉 450 nm). By combining fluorescence profiles with chlorophyll distribution across the leaf, the profile of the apparent extinction coefficient was calculated. The light profiles within spinach leaves could be well approximated by an apparent extinction coefficient and the Beer–Lambert/Bouguer laws. Light was absorbed at greater depths than predicted from fibre optic measurements, with 50% of blue and green light reaching 125 and 240 µm deep, respectively.

Notizen: Abstract. Light gradients were measured and correlated with chlorophyll concentration and anatomy of leaves in spinach (Spinacia oleracea L.). Light gradients were measured at 450, 550 and 680 nm within thin (455 μm) and thick (630 μm) leaves of spinach grown under sun and shade conditions. The light gradients were relatively steep in both types of leaves and 90% of the light at 450 and 680 nm was absorbed by the initial 140 μm of the palisade. In general, blue light was depleted faster than red light which, in turn was depleted faster than green light. Light penetrated further into the thicker palisade of sun leaves in comparison to the shade leaves. The distance that blue light at 450 nm travelled before it became 90% depleted was 120 μm in sun leaves versus 76 μm in shade leaves. Red light at 680 nm and green light at 550 nm travelled further but the trends were similar to that measured at 450nm. The steeper light gradients within the palisade-of shade leaves were caused by increased scattering of light within the intercellular air spaces and/or cells which were less compact than those in sun leaves. The decline in the amount of light within the leaf appeared to be balanced by a gradient in chlorophyll concentration measured in paradermal sections. Progressing from the adaxial epidermis, chlorophyll content increased through the palisade and then declined through the spongy mesophyll. Chlorophyll content was similar in the palisade of both sun and shade leaves. Chloroplast distribution within both sun and shade leaves was relatively uniform so that the chlorophyll gradient appeared to be caused by greater amounts of chlorophyll within chloroplasts located deeper within the leaf. These results indicate that the anatomy of the palisade may be of special importance for controlling the penetration of photo-synthetically active radiation into the leaf. Changing the structural characteristics of individual palisade cells or their arrangement may be an adaptation that maximizes the absorption of light in leaves with varying mesophyll thickness due to different ambient light regimes.

Notizen: Profiles of chlorophyll fluorescence were measured in spinach leaves irradiated with monochromatic light. The characteristics of the profiles within the mesophyll were determined by the optical properties of the leaf tissue and the spectral quality of the actinic light. When leaves were infiltrated with 10−4M DCMU [3-(3,4-dichlorophenyl)-1, 1-dimethyl-urea] or water, treatments that minimized light scattering, irradiation with 2000 μmol m−2 s−1 green light produced broad Gaussian-shaped fluorescence profiles that spanned most of the mesophyll. Profiles for chlorophyll fluorescence in the red (680 ± 16 nm) and far red (λ 〉 710 nm) were similar except that there was elevated red fluorescence near the adaxial leaf surface relative to far red fluorescence. Fluorescence profiles were narrower in non-infiltrated leaf samples where light scattering increased the light gradient. The fluorescence profile was broader when the leaf was irradiated on its adaxial versus abaxial surface due to the contrasting optical properties of the palisade and spongy mesophyll. Irradiation with blue, red and green monochromatic light produced profiles that peaked 50, 100 and 150 μm, respectively, beneath the irradiated surface. These results are consistent with previous measurements of the light gradient in spinach and they agree qualitatively with measurements of carbon fixation under monochromatic blue, red and green light. These results suggest that chlorophyll fluorescence profiles may be used to estimate the distribution of quanta that are absorbed within the leaf for photosynthesis.

Notizen: In some plants, particularly herbaceous species, a considerable proportion of incident ultraviolet-B radiation (UV-B, 280-320 nm) penetrates into the leaf mesophyll where it is potentially damaging to nucleic acids and the photosyn-thetic machinery. We used optical techniques to look at the spatial variation in UV-B penetration through the epidermis of foliage of two herbaceous species (Chenopodium album and Smilacina stellata)and a conifer (Picea pun-gens). Measurements of UV-B penetration in intact foliage with a fibre-optic microprobe revealed that 300 nm radiation reached 161±36μm (mean±SD) into leaves of C. album, 154±40μm in S. stellata and 17±2μm in P. pungens, with epidermal transmittance being 39±14%, 55±19% and 0%, respectively. A thin polymer film was developed which fluoresced blue when irradiated by UV-B. Fresh epidermal leaf peels were placed over the film and irradiated with UV-B, and microscopic examination of the film from below allowed us to determine the spatial pattern of UV-B penetration through the epidermis. In herbaceous species, film fluorescence below cell walls, but not epidermal and guard cell protoplasts indicated that UV-B transmittance was much greater through anticlinal cell wall regions than protoplasts. Ultraviolet-B transmittance through large areas of epidermal cells could be induced by plasmolysis. Epidermal transmittance was also relatively high through stomal pores (and what appear to be nuclei in Smilacina), but relatively low through stomatal guard cells. Results from the fluorescing film technique were substantiated by direct measurements of UV-B transmittance through epidermal peels with a fibre-optic microprobe run paradermally along the bottom or inner side of irradiated peels. In Smilacina, we estimate that UV-B epidermal transmittance was up to 90% through anticlinal cell wall regions, but 〈10% through protoplast areas. In contrast to herbaceous species, we did not detect any UV-B transmittance through the epidermis of P. pungens with either the fluorescing film or the fibre-optic microprobe technique. The epidermis appears to be a much more spatially uniform UV-B filter in conifers than in these herbaceous species.

Notizen: Light gradients were measured in leaves that had different types of anatomical development of the mesophyll but similar pigment content. Leaves of the legume, Thermopsis montana, had columnar palisade and spongy mesophyll whereas leaves of the monocot, Smilacina stellata, had spongy mesophyll only. Light gradients were measured at 550 nm in both types of leaves when they were irradiated with collimated or diffuse light. When irradiated with collimated light, light gradients were steeper in leaves with spongy mesophyll in comparison to those that had palisade tissue. On the other hand, light gradients were similar between both leaf types when they were irradiated with diffuse light. Thus, columnar palisade cells facilitated the penetration of collimated light over diffuse light. These results suggest that palisade tissue may help distribute light more uniformly to chloroplasts within the leaf. Moreover, the functional significance of palisade tissue may be related to the amount of collimated light within the natural environment.

Notizen: Abstract. The distribution of chlorophyll fluorescence was measured within leaves of Medicago saliva with a fibre optic microprobe. Leaves were irradiated with broad band blue light (1000 μmol m−2s−1) and chlorophyll fluorescence was measured at 688 nm. The amount of fluorescence measured within the leaf depended upon the direction in which the probe was inserted. When the probe was advanced directly through the leaf from the shaded towards the irradiated surface, the maximum amount of detected fluorescence occurred near the boundary between the palisade and spongy mesophyll. When the probe was advanced through the leaf from the opposite direction maximum detected fluorescence was at the boundary between the epidermis and palisade. These results appear to be a consequence of the blue light gradient, which declined exponentially within the palisade but was counterbalanced by increasing chlorophyll content within the leaf. Modelling indicates that the measured distribution of chlorophyll fluorescence can be explained by relatively uniform emission of fluorescence throughout the palisade layer, indicating that the chloroplasts may be photosynthetically specialized to their light environment within the leaf.

Notizen: Abstract. In Oxalis, epidermal cells on both the adaxial and abaxial surface of the leaf concentrated light within the leaf by a lens mechanism. Focal lengths of epidermal cells were estimated using two methods: they were calculated from radius of curvature measurements taken from individual epidermal cells, and were measured directly in agarose replicas of the leaf surface. In the three species of Oxalis examined, light that was incident upon the adaxial leaf surface was concentrated within the palisade, whereas light that was incident upon the abaxial leaf surface was concentrated within the spongy mesophyll. Using sensiometric analysis, theoretically maximal focal intesifications were measured in leaf replicas at the focal maximum and at intermediate positions corresponding to the mid-region of the palisade and spongy mesophyll tissues. Focal intensifications ranged from 2.2 to 10.4 times incident light at the focal maximum, and 1.3 to 4.5 in the palisade or spongy mesophyll layers. Elimination of epidermal focussing, by covering the leaf surface with a thin layer of mineral oil, strongly affected chlorophyll fluorescence induction curves resulting in a decrease of 10–40% in the initial (F0) and variable fluorescence (Fv). These results are consistent with the interpretation that the chloroplasts were adapted to their light microenvironment within the leaf and that focussing by the epidermis channelled light to a population of chloroplasts that were adapted to high light.