The intensification of absorbance changes in leaves by light-dispersion

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Abstract
In dispersive samples, like leaves, the absorbance of pigments is intensified. The intensification is due to a longer optical path through the dispersive sample. However, in chloroplast suspensions the optical path is not much longer than in clear solutions. The factor of intensification β (=the lengthening of the optical path) is calculated by comparing the absorbance of leaves and the absorbance of chloroplast suspensions with equal pigment-content. This method also includes the influence of possible sieve effects which could decrease absorbance. The measurements are carried out with high- and low-light leaves of different thickness and pigment content. The intensification of absorbance was 2–2.5 fold. In highlight leaves it was somewhat less than in low-light leaves. The factor β is better correlated to the pigment content than to the thickness of the leaves. The plot of absorbance versus the pigment content of the leaves shows that β decreases with increasing pigment content. In contrast, chloroplast suspensions show a linear dependence as expected from Lambert-Beer's law. Thus, in leaves with very low pigment content the absorbance is intensified up to 6 fold while the intensification decreases with increasing absorbance. These results are in good agreement with measurements of Tsel'niker (1975) and with the theoretical predictions of Butler's formula (1960). Absorbance changes due to photooxidation of P-700 and cytochrome f in intact leaves are measured, and β is used to calculate the amount of the oxidized components. Without correction for β the values would be much greater than the amount actually present. The corrected data show that between 70 and 90% of the present P-700 and cytochrome f can be photooxidized in the intact leaf.