Measurement of the optical properties of leaves under diffuse light.

Measuring leaf light absorptance is central to many areas of plant biology including photosynthesis and energy balance. Absorptance is calculated from measured values of transmittance and reflectance, and most such measurements have used direct beam light. However, photosynthesis and other processes can differ under direct and diffuse light. Optical properties under diffuse light may be different, but there have been technical difficulties involved in measuring total reflectance of diffuse light. We developed instrumentation to measure this reflectance using a chopped measuring beam delivered alternately to sample and reference integrating spheres, and lock-in detection. We also built instrumentation for measuring transmittance of diffuse light. We developed standards to calibrate our instruments and correct for substitution error, a known systematic error with integrating sphere-based measurements. Helianthus annuus leaves measured under diffuse light reflected 5-10% more and transmitted a few percent less 400-700 nm light than under direct light. Overall absorptance was only a few percent higher under direct light, but leaves may utilize absorbed direct and diffuse light differently. For example, of the light entering the leaf, significantly more direct light than diffuse light is transmitted through the leaf, suggesting differences in localization of absorption within the leaf.

A new paradigm in leaf-level photosynthesis: direct and diffuse lights are not equal.

Global-change scenarios suggest a trend of increasing diffuse light due to expected increases in cloud cover. Canopy-level measurements of plant-community photosynthesis under diffuse light show increased productivity attributed to more uniform distribution of light within the forest canopy, yet the effect of the directional quality of light at the leaf level is unknown. Here we show that leaf-level photosynthesis in sun leaves of both C(3) and C(4) plants can be 10-15% higher under direct light compared to equivalent absorbed irradiances of diffuse light. High-light-grown leaves showed significant photosynthetic enhancement in direct light, while shade-adapted leaves showed no preference for direct or diffuse light at any irradiance. High-light-grown leaves with multiple palisade layers may be adapted to better utilize direct than diffuse light, while shade leaf structure does not appear to discriminate light based on its directionality. Based upon our measurements, it appears that leaf-level and canopy-level photosynthetic processes react differently to the directionality of light, and previously observed increases in canopy-level photosynthesis occur even though leaf-level photosynthesis decreases under diffuse light.

Photoacoustic analysis indicates that chloroplast movement does not alter liquid-phase CO2 diffusion in leaves of Alocasia brisbanensis.

Light-mediated chloroplast movements are common in plants. When leaves of Alocasia brisbanensis (F.M. Bailey) Domin are exposed to dim light, mesophyll chloroplasts spread along the periclinal walls normal to the light, maximizing absorbance. Under high light, the chloroplasts move to anticlinal walls. It has been proposed that movement to the high-light position shortens the diffusion path for CO(2) from the intercellular air spaces to the chloroplasts, thus reducing CO(2) limitation of photosynthesis. To test this hypothesis, we used pulsed photoacoustics to measure oxygen diffusion times as a proxy for CO(2) diffusion in leaf cells. We found no evidence that chloroplast movement to the high-light position enhanced gas diffusion. Times for oxygen diffusion were not shorter in leaves pretreated with white light, which induced chloroplast movement to the high-light position, compared with leaves pretreated with 500 to 700 nm light, which did not induce movement. From the oxygen diffusion time and the diffusion distance from chloroplasts to the intercellular gas space, we calculated an oxygen permeability of 2.25 x 10(-)(6) cm(2) s(-)(1) for leaf cells at 20 degrees C. When leaf temperature was varied from 5 degrees C to 40 degrees C, the permeability for oxygen increased between 5 degrees C and 20 degrees C but changed little between 20 degrees C and 40 degrees C, indicating changes in viscosity or other physical parameters of leaf cells above 20 degrees C. Resistance for CO(2) estimated from oxygen permeability was in good agreement with published values, validating photoacoustics as another way of assessing internal resistances to CO(2) diffusion.

Ultraviolet radiation and the snow alga Chlamydomonas nivalis (Bauer) Wille.

Aplanospores of Chlamydomonas nivalis are frequently found in high-altitude, persistent snowfields where they are photosynthetically active despite cold temperatures and high levels of visible and ultraviolet (UV) radiation. The goals of this work were to characterize the UV environment of the cells in the snow and to investigate the existence and localization of screening compounds that might prevent UV damage. UV irradiance decreased precipitously in snow, with UV radiation of wavelengths 280-315 nm and UV radiation of wavelengths 315-400 nm dropping to 50% of incident levels in the top 1 and 2 cm, respectively. Isolated cell walls exhibited UV absorbance, possibly by sporopollenin, but this absorbance was weak in images of broken or plasmolyzed cells observed through a UV microscope. The cells also contained UV-absorbing cytoplasmic compounds, with the extrachloroplastic carotenoid astaxanthin providing most of the screening. Additional screening compound(s) soluble in aqueous methanol with an absorption maximum at 335 nm played a minor role. Thus, cells are protected against potentially high levels of UV radiation by the snow itself when they live several centimeters beneath the surface, and they rely on cellular screening compounds, chiefly astaxanthin, when located near the surface where UV fluxes are high.

Surface gas-exchange processes of snow algae.

The red-colored chlorophyte Chlamydomonas nivalis is commonly found in summer snowfields. We used a modified Li-Cor gas-exchange system to investigate surface gas-exchange characteristics of snow colonized by this alga, finding rates of CO(2) uptake up to 0.3 micromol.m(-2).s(-1) in dense algal blooms. Experiments varying the irradiance resulted in light curves that resembled those of the leaves of higher plants. Red light was more effective than white and much more effective than green or blue, because of the red astaxanthin that surrounds and masks the algal chloroplasts. Integrating daily course measurements of gas exchange showed CO(2) uptake around 2,300 micromol.m(-2).day(-1) in heavily colonized patches, indicating that summer snowfields can be surprisingly productive.