Institut für Allgemeine Botanik der Universität Mainz, Saarstraße 21, D-6500, Mainz, Federal Republic of Germany.
Planta. 1979 Oct;146(5):551-7. doi: 10.1007/BF00388831.
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.
在分散的样品中,如叶片,色素的吸光度会增强。这种增强是由于分散样品的光程更长。然而,在叶绿体悬浮液中,光程并不比澄清溶液长很多。增强因子β(=光程的延长)是通过比较具有相同色素含量的叶片和叶绿体悬浮液的吸光度来计算的。这种方法还包括可能降低吸光度的筛效应的影响。测量是用不同厚度和色素含量的高光和低光叶片进行的。吸光度的增强为 2-2.5 倍。在高光叶片中,它比在低光叶片中略小。β因子与色素含量的相关性优于与叶片厚度的相关性。吸光度与叶片色素含量的关系图表明,β随色素含量的增加而减小。相比之下,叶绿体悬浮液显示出与朗伯-比尔定律预期的线性关系。因此,在色素含量非常低的叶片中,吸光度增强了 6 倍,而随着吸光度的增加,增强程度降低。这些结果与 Tsel'niker(1975)的测量结果以及 Butler 公式(1960)的理论预测非常吻合。测量了完整叶片中 P-700 和细胞色素 f 的光氧化引起的吸光度变化,并使用β来计算氧化组分的量。如果不进行β校正,这些值将比实际存在的量大得多。经校正的数据表明,在完整叶片中,目前 P-700 和细胞色素 f 的 70%至 90%可以被光氧化。